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Special Issue "Protein Crystallography in Molecular Biology"

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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Biochemistry, Molecular Biology and Biophysics".

Deadline for manuscript submissions: closed (31 May 2012)

Special Issue Editor

Guest Editor
Prof. Dr. Charles A. Collyer

School of Molecular Bioscience, G08 - Biochemistry Building, The University of Sydney, NSW 2006, Australia
Website | E-Mail
Interests: protein structure and function; anaerobic bacteria; bacterial adhesins

Special Issue Information

Dear Colleagues,

Our understanding of how biological molecules interact is often presented as a number of snapshots of static images derived from bio-molecular crystallography (and today increasingly derived also from NMR spectroscopy). We “see” our molecular biology in these images as processes of recognition, biochemical transformation, physical movement and communication. Being an imaging technique it provides unbiased insights into the complex features of proteins, DNA, RNA, and their relationship to substrates, inhibitors and binding partners. Crystal structures of proteins often surprise as they produce images of molecules as they really are! Many novel gene products are not structured how we initially perceive them to be but experimentally determined structures enable us to accurately delineate the genetic information into three dimensions. By identifying particular structural features of biological molecules with specific functions biologists can be led to focus on mechanistic relationships and then to create working models that can usefully drive studies of complex systems in biology. This type of approach using the identification of key elements of structure to start this discovery process has had application in all of biology, from genetics to cell biology and then beyond into our understanding of evolution.

This special issue on protein crystallography for the International Journal of Molecular Sciences will focus on examples in molecular biology where structural data derived from protein crystallography has “turned heads” and then provided the means to generate unique insights into molecular processes in biology.

Prof. Dr. Charles A. Collyer
Guest Editor

Keywords

  • Sub-domain structure
  • structural motifs
  • protein families
  • protein/nucleic acid recognition
  • structural model
  • conformational signalling
  • enzyme mechanism
  • activation
  • inhibition
  • regulation

Related Special Issue

Published Papers (15 papers)

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Research

Jump to: Review

Open AccessCommunication Proteins of Unknown Function in the Protein Data Bank (PDB): An Inventory of True Uncharacterized Proteins and Computational Tools for Their Analysis
Int. J. Mol. Sci. 2012, 13(10), 12761-12772; doi:10.3390/ijms131012761
Received: 11 September 2012 / Revised: 21 September 2012 / Accepted: 21 September 2012 / Published: 8 October 2012
Cited by 10 | PDF Full-text (664 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
Proteins of uncharacterized functions form a large part of many of the currently available biological databases and this situation exists even in the Protein Data Bank (PDB). Our analysis of recent PDB data revealed that only 42.53% of PDB entries (1084 coordinate files)
[...] Read more.
Proteins of uncharacterized functions form a large part of many of the currently available biological databases and this situation exists even in the Protein Data Bank (PDB). Our analysis of recent PDB data revealed that only 42.53% of PDB entries (1084 coordinate files) that were categorized under “unknown function” are true examples of proteins of unknown function at this point in time. The remainder 1465 entries also annotated as such appear to be able to have their annotations re-assessed, based on the availability of direct functional characterization experiments for the protein itself, or for homologous sequences or structures thus enabling computational function inference. Full article
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology)
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Open AccessArticle Direct Observation of Protein Microcrystals in Crystallization Buffer by Atmospheric Scanning Electron Microscopy
Int. J. Mol. Sci. 2012, 13(8), 10553-10567; doi:10.3390/ijms130810553
Received: 1 June 2012 / Revised: 2 August 2012 / Accepted: 3 August 2012 / Published: 22 August 2012
Cited by 12 | PDF Full-text (4962 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
X-ray crystallography requires high quality crystals above a given size. This requirement not only limits the proteins to be analyzed, but also reduces the speed of the structure determination. Indeed, the tertiary structures of many physiologically important proteins remain elusive because of the
[...] Read more.
X-ray crystallography requires high quality crystals above a given size. This requirement not only limits the proteins to be analyzed, but also reduces the speed of the structure determination. Indeed, the tertiary structures of many physiologically important proteins remain elusive because of the so-called “crystallization bottleneck”. Once microcrystals have been obtained, crystallization conditions can be optimized to produce bigger and better crystals. However, the identification of microcrystals can be difficult due to the resolution limit of optical microscopy. Electron microscopy has sometimes been utilized instead, with the disadvantage that the microcrystals usually must be observed in vacuum, which precludes the usage for crystal screening. The atmospheric scanning electron microscope (ASEM) allows samples to be observed in solution. Here, we report the use of this instrument in combination with a special thin-membrane dish with a crystallization well. It was possible to observe protein crystals of lysozyme, lipase B and a histone chaperone TAF-Iβ in crystallization buffers, without the use of staining procedures. The smallest crystals observed with ASEM were a few µm in width, and ASEM can be used with non-transparent solutions. Furthermore, the growth of salt crystals could be monitored in the ASEM, and the difference in contrast between salt and protein crystals made it easy to distinguish between these two types of microcrystals. These results indicate that the ASEM could be an important new tool for the screening of protein microcrystals. Full article
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology)
Figures

Open AccessArticle Effects of a Buried Cysteine-To-Serine Mutation on Yeast Triosephosphate Isomerase Structure and Stability
Int. J. Mol. Sci. 2012, 13(8), 10010-10021; doi:10.3390/ijms130810010
Received: 4 June 2012 / Revised: 24 July 2012 / Accepted: 26 July 2012 / Published: 10 August 2012
Cited by 3 | PDF Full-text (766 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
All the members of the triosephosphate isomerase (TIM) family possess a cystein residue (Cys126) located near the catalytically essential Glu165. The evolutionarily conserved Cys126, however, does not seem to play a significant role in the catalytic activity. On the other hand, substitution of
[...] Read more.
All the members of the triosephosphate isomerase (TIM) family possess a cystein residue (Cys126) located near the catalytically essential Glu165. The evolutionarily conserved Cys126, however, does not seem to play a significant role in the catalytic activity. On the other hand, substitution of this residue by other amino acid residues destabilizes the dimeric enzyme, especially when Cys is replaced by Ser. In trying to assess the origin of this destabilization we have determined the crystal structure of Saccharomyces cerevisiae TIM (ScTIM) at 1.86 Å resolution in the presence of PGA, which is only bound to one subunit. Comparisons of the wild type and mutant structures reveal that a change in the orientation of the Ser hydroxyl group, with respect to the Cys sulfhydryl group, leads to penetration of water molecules and apparent destabilization of residues 132–138. The latter results were confirmed by means of Molecular Dynamics, which showed that this region, in the mutated enzyme, collapses at about 70 ns. Full article
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology)
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Open AccessArticle Correlation between Protein Sequence Similarity and Crystallization Reagents in the Biological Macromolecule Crystallization Database
Int. J. Mol. Sci. 2012, 13(8), 9514-9526; doi:10.3390/ijms13089514
Received: 1 June 2012 / Revised: 9 July 2012 / Accepted: 10 July 2012 / Published: 27 July 2012
Cited by 6 | PDF Full-text (469 KB) | HTML Full-text | XML Full-text
Abstract
The protein structural entries grew far slower than the sequence entries. This is partly due to the bottleneck in obtaining diffraction quality protein crystals for structural determination using X-ray crystallography. The first step to achieve protein crystallization is to find out suitable chemical
[...] Read more.
The protein structural entries grew far slower than the sequence entries. This is partly due to the bottleneck in obtaining diffraction quality protein crystals for structural determination using X-ray crystallography. The first step to achieve protein crystallization is to find out suitable chemical reagents. However, it is not an easy task. Exhausting trial and error tests of numerous combinations of different reagents mixed with the protein solution are usually necessary to screen out the pursuing crystallization conditions. Therefore, any attempts to help find suitable reagents for protein crystallization are helpful. In this paper, an analysis of the relationship between the protein sequence similarity and the crystallization reagents according to the information from the existing databases is presented. We extracted information of reagents and sequences from the Biological Macromolecule Crystallization Database (BMCD) and the Protein Data Bank (PDB) database, classified the proteins into different clusters according to the sequence similarity, and statistically analyzed the relationship between the sequence similarity and the crystallization reagents. The results showed that there is a pronounced positive correlation between them. Therefore, according to the correlation, prediction of feasible chemical reagents that are suitable to be used in crystallization screens for a specific protein is possible. Full article
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology)
Open AccessArticle 3D Structure Elucidation of Thermostable L2 Lipase from Thermophilic Bacillus sp. L2
Int. J. Mol. Sci. 2012, 13(7), 9207-9217; doi:10.3390/ijms13079207
Received: 16 May 2012 / Revised: 29 June 2012 / Accepted: 12 July 2012 / Published: 23 July 2012
Cited by 2 | PDF Full-text (1776 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
The crystallization of proteins makes it possible to determine their structure by X-ray crystallography, and is therefore important for the analysis of protein structure-function relationships. L2 lipase was crystallized by using the J-tube counter diffusion method. A crystallization consisting of 20% PEG 6000,
[...] Read more.
The crystallization of proteins makes it possible to determine their structure by X-ray crystallography, and is therefore important for the analysis of protein structure-function relationships. L2 lipase was crystallized by using the J-tube counter diffusion method. A crystallization consisting of 20% PEG 6000, 50 mM MES pH 6.5 and 50 mM NaCl was found to be the best condition to produce crystals with good shape and size (0.5 × 0.1 × 0.2 mm). The protein concentration used for the crystallization was 3 mg/mL. L2 lipase crystal has two crystal forms, Shape 1 and Shape 2. Shape 2 L2 lipase crystal was diffracted at 1.5 Å and the crystal belongs to the orthorhombic space group P212121, with unit-cell parameters a = 72.0, b = 81.8, c = 83.4 Å, α = β = γ = 90°. There is one molecule per asymmetric unit and the solvent content of the crystals is 56.9%, with a Matthew’s coefficient of 2.85 Å Da−1. The 3D structure of L2 lipase revealed topological organization of α/β-hydrolase fold consisting of 11 β-strands and 13 α-helices. Ser-113, His-358 and Asp-317 were assigned as catalytic triad residues. One Ca2+ and one Zn2+ were found in the L2 lipase molecule. Full article
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology)
Open AccessArticle Structural Analysis of Cytochrome P450 105N1 Involved in the Biosynthesis of the Zincophore, Coelibactin
Int. J. Mol. Sci. 2012, 13(7), 8500-8513; doi:10.3390/ijms13078500
Received: 1 June 2012 / Revised: 22 June 2012 / Accepted: 28 June 2012 / Published: 9 July 2012
Cited by 11 | PDF Full-text (375 KB) | HTML Full-text | XML Full-text
Abstract
Coelibactin is a putative non-ribosomally synthesized peptide with predicted zincophore activity and which has been implicated in antibiotic regulation in Streptomyces coelicolor A3(2). The coelibactin biosynthetic pathway contains a stereo- and regio-specific monooxygenation step catalyzed by a cytochrome P450 enzyme (CYP105N1). We have
[...] Read more.
Coelibactin is a putative non-ribosomally synthesized peptide with predicted zincophore activity and which has been implicated in antibiotic regulation in Streptomyces coelicolor A3(2). The coelibactin biosynthetic pathway contains a stereo- and regio-specific monooxygenation step catalyzed by a cytochrome P450 enzyme (CYP105N1). We have determined the X-ray crystal structure of CYP105N1 at 2.9 Å and analyzed it in the context of the bacterial CYP105 family as a whole. The crystal structure reveals a channel between the α-helical domain and the β-sheet domain exposing the heme pocket and the long helix I to the solvent. This wide-open conformation of CYP105N1 may be related to the bulky substrate coelibactin. The ligand-free CYP105N1 structure has enough room in the substrate access channel to allow the coelibactin to enter into the active site. Analysis of typical siderophore ligands suggests that CYP105N1 may produce derivatives of coelibactin, which would then be able to chelate the zinc divalent cation. Full article
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology)
Open AccessArticle High Resolution Crystal Structures of the Cerebratulus lacteus Mini-Hb in the Unligated and Carbomonoxy States
Int. J. Mol. Sci. 2012, 13(7), 8025-8037; doi:10.3390/ijms13078025
Received: 31 May 2012 / Revised: 14 June 2012 / Accepted: 15 June 2012 / Published: 28 June 2012
Cited by 2 | PDF Full-text (746 KB) | HTML Full-text | XML Full-text
Abstract
The nerve tissue mini-hemoglobin from Cerebratulus lacteus (CerHb) displays an essential globinfold hosting a protein matrix tunnel held to allow traffic of small ligands to and from the heme. CerHb heme pocket hosts the distal TyrB10/GlnE7 pair, normally linked to low rates
[...] Read more.
The nerve tissue mini-hemoglobin from Cerebratulus lacteus (CerHb) displays an essential globin fold hosting a protein matrix tunnel held to allow traffic of small ligands to and from the heme. CerHb heme pocket hosts the distal TyrB10/GlnE7 pair, normally linked to low rates of O2 dissociation and ultra-high O2 affinity. However, CerHb affinity for O2 is similar to that of mammalian myoglobins, due to a dynamic equilibrium between high and low affinity states driven by the ability of ThrE11 to orient the TyrB10 OH group relative to the heme ligand. We present here the high resolution crystal structures of CerHb in the unligated and carbomonoxy states. Although CO binds to the heme with an orientation different from the O2 ligand, the overall binding schemes for CO and O2 are essentially the same, both ligands being stabilized through a network of hydrogen bonds based on TyrB10, GlnE7, and ThrE11. No dramatic protein structural changes are needed to support binding of the ligands, which can freely reach the heme distal site through the apolar tunnel. A lack of main conformational changes between the heme-unligated and -ligated states grants stability to the folded mini-Hb and is a prerequisite for fast ligand diffusion to/from the heme. Full article
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology)
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Open AccessArticle Improvement of Thermal Stability via Outer-Loop Ion Pair Interaction of Mutated T1 Lipase from Geobacillus zalihae Strain T1
Int. J. Mol. Sci. 2012, 13(1), 943-960; doi:10.3390/ijms13010943
Received: 8 October 2011 / Revised: 25 November 2011 / Accepted: 28 November 2011 / Published: 17 January 2012
Cited by 13 | PDF Full-text (1631 KB) | HTML Full-text | XML Full-text
Abstract
Mutant D311E and K344R were constructed using site-directed mutagenesis to introduce an additional ion pair at the inter-loop and the intra-loop, respectively, to determine the effect of ion pairs on the stability of T1 lipase isolated from Geobacillus zalihae. A series of
[...] Read more.
Mutant D311E and K344R were constructed using site-directed mutagenesis to introduce an additional ion pair at the inter-loop and the intra-loop, respectively, to determine the effect of ion pairs on the stability of T1 lipase isolated from Geobacillus zalihae. A series of purification steps was applied, and the pure lipases of T1, D311E and K344R were obtained. The wild-type and mutant lipases were analyzed using circular dichroism. The Tm for T1 lipase, D311E lipase and K344R lipase were approximately 68.52 °C, 70.59 °C and 68.54 °C, respectively. Mutation at D311 increases the stability of T1 lipase and exhibited higher Tm as compared to the wild-type and K344R. Based on the above, D311E lipase was chosen for further study. D311E lipase was successfully crystallized using the sitting drop vapor diffusion method. The crystal was diffracted at 2.1 Å using an in-house X-ray beam and belonged to the monoclinic space group C2 with the unit cell parameters a = 117.32 Å, b = 81.16 Å and c = 100.14 Å. Structural analysis showed the existence of an additional ion pair around E311 in the structure of D311E. The additional ion pair in D311E may regulate the stability of this mutant lipase at high temperatures as predicted in silico and spectroscopically. Full article
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology)

Review

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Open AccessReview Fragment-Based Screening by Protein Crystallography: Successes and Pitfalls
Int. J. Mol. Sci. 2012, 13(10), 12857-12879; doi:10.3390/ijms131012857
Received: 4 July 2012 / Revised: 30 August 2012 / Accepted: 19 September 2012 / Published: 8 October 2012
Cited by 14 | PDF Full-text (802 KB) | HTML Full-text | XML Full-text
Abstract
Fragment-based drug discovery (FBDD) concerns the screening of low-molecular weight compounds against macromolecular targets of clinical relevance. These compounds act as starting points for the development of drugs. FBDD has evolved and grown in popularity over the past 15 years. In this paper,
[...] Read more.
Fragment-based drug discovery (FBDD) concerns the screening of low-molecular weight compounds against macromolecular targets of clinical relevance. These compounds act as starting points for the development of drugs. FBDD has evolved and grown in popularity over the past 15 years. In this paper, the rationale and technology behind the use of X-ray crystallography in fragment based screening (FBS) will be described, including fragment library design and use of synchrotron radiation and robotics for high-throughput X-ray data collection. Some recent uses of crystallography in FBS will be described in detail, including interrogation of the drug targets β-secretase, phenylethanolamine N-methyltransferase, phosphodiesterase 4A and Hsp90. These examples provide illustrations of projects where crystallography is straightforward or difficult, and where other screening methods can help overcome the limitations of crystallography necessitated by diffraction quality. Full article
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology)
Open AccessReview Characterization of Aptamer-Protein Complexes by X-ray Crystallography and Alternative Approaches
Int. J. Mol. Sci. 2012, 13(8), 10537-10552; doi:10.3390/ijms130810537
Received: 31 May 2012 / Revised: 9 August 2012 / Accepted: 17 August 2012 / Published: 22 August 2012
Cited by 13 | PDF Full-text (1413 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
Aptamers are oligonucleotide ligands, either RNA or ssDNA, selected for high-affinity binding to molecular targets, such as small organic molecules, proteins or whole microorganisms. While reports of new aptamers are numerous, characterization of their specific interaction is often restricted to the affinity of
[...] Read more.
Aptamers are oligonucleotide ligands, either RNA or ssDNA, selected for high-affinity binding to molecular targets, such as small organic molecules, proteins or whole microorganisms. While reports of new aptamers are numerous, characterization of their specific interaction is often restricted to the affinity of binding (KD). Over the years, crystal structures of aptamer-protein complexes have only scarcely become available. Here we describe some relevant technical issues about the process of crystallizing aptamer-protein complexes and highlight some biochemical details on the molecular basis of selected aptamer-protein interactions. In addition, alternative experimental and computational approaches are discussed to study aptamer-protein interactions. Full article
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology)
Open AccessReview Biogenesis and Mechanism of Action of Small Non-Coding RNAs: Insights from the Point of View of Structural Biology
Int. J. Mol. Sci. 2012, 13(8), 10268-10295; doi:10.3390/ijms130810268
Received: 30 May 2012 / Revised: 17 July 2012 / Accepted: 2 August 2012 / Published: 17 August 2012
Cited by 3 | PDF Full-text (3289 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
Non-coding RNAs are dominant in the genomic output of the higher organisms being not simply occasional transcripts with idiosyncratic functions, but constituting an extensive regulatory network. Among all the species of non-coding RNAs, small non-coding RNAs (miRNAs, siRNAs and piRNAs) have been shown
[...] Read more.
Non-coding RNAs are dominant in the genomic output of the higher organisms being not simply occasional transcripts with idiosyncratic functions, but constituting an extensive regulatory network. Among all the species of non-coding RNAs, small non-coding RNAs (miRNAs, siRNAs and piRNAs) have been shown to be in the core of the regulatory machinery of all the genomic output in eukaryotic cells. Small non-coding RNAs are produced by several pathways containing specialized enzymes that process RNA transcripts. The mechanism of action of these molecules is also ensured by a group of effector proteins that are commonly engaged within high molecular weight protein-RNA complexes. In the last decade, the contribution of structural biology has been essential to the dissection of the molecular mechanisms involved in the biosynthesis and function of small non-coding RNAs. Full article
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology)
Open AccessReview Function and 3D Structure of the N-Glycans on Glycoproteins
Int. J. Mol. Sci. 2012, 13(7), 8398-8429; doi:10.3390/ijms13078398
Received: 31 May 2012 / Revised: 18 June 2012 / Accepted: 28 June 2012 / Published: 6 July 2012
Cited by 27 | PDF Full-text (3679 KB) | HTML Full-text | XML Full-text
Abstract
Glycosylation is one of the most common post-translational modifications in eukaryotic cells and plays important roles in many biological processes, such as the immune response and protein quality control systems. It has been notoriously difficult to study glycoproteins by X-ray crystallography since the
[...] Read more.
Glycosylation is one of the most common post-translational modifications in eukaryotic cells and plays important roles in many biological processes, such as the immune response and protein quality control systems. It has been notoriously difficult to study glycoproteins by X-ray crystallography since the glycan moieties usually have a heterogeneous chemical structure and conformation, and are often mobile. Nonetheless, recent technical advances in glycoprotein crystallography have accelerated the accumulation of 3D structural information. Statistical analysis of “snapshots” of glycoproteins can provide clues to understanding their structural and dynamic aspects. In this review, we provide an overview of crystallographic analyses of glycoproteins, in which electron density of the glycan moiety is clearly observed. These well-defined N-glycan structures are in most cases attributed to carbohydrate-protein and/or carbohydrate-carbohydrate interactions and may function as “molecular glue” to help stabilize inter- and intra-molecular interactions. However, the more mobile N-glycans on cell surface receptors, the electron density of which is usually missing on X-ray crystallography, seem to guide the partner ligand to its binding site and prevent irregular protein aggregation by covering oligomerization sites away from the ligand-binding site. Full article
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology)
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Open AccessReview 8-Oxoguanine DNA Glycosylases: One Lesion, Three Subfamilies
Int. J. Mol. Sci. 2012, 13(6), 6711-6729; doi:10.3390/ijms13066711
Received: 20 April 2012 / Revised: 14 May 2012 / Accepted: 24 May 2012 / Published: 1 June 2012
Cited by 8 | PDF Full-text (2831 KB) | HTML Full-text | XML Full-text
Abstract
Amongst the four bases that form DNA, guanine is the most susceptible to oxidation, and its oxidation product, 7,8-dihydro-8-oxoguanine (8-oxoG) is the most prevalent base lesion found in DNA. Fortunately, throughout evolution cells have developed repair mechanisms, such as the 8-oxoguanine DNA glycosylases
[...] Read more.
Amongst the four bases that form DNA, guanine is the most susceptible to oxidation, and its oxidation product, 7,8-dihydro-8-oxoguanine (8-oxoG) is the most prevalent base lesion found in DNA. Fortunately, throughout evolution cells have developed repair mechanisms, such as the 8-oxoguanine DNA glycosylases (OGG), which recognize and excise 8-oxoG from DNA thereby preventing the accumulation of deleterious mutations. OGG are divided into three subfamilies, OGG1, OGG2 and AGOG, which are all involved in the base excision repair (BER) pathway. The published structures of OGG1 and AGOG, as well as the recent availability of OGG2 structures in both apo- and liganded forms, provide an excellent opportunity to compare the structural and functional properties of the three OGG subfamilies. Among the observed differences, the three-dimensional fold varies considerably between OGG1 and OGG2 members, as the latter lack the A-domain involved in 8-oxoG binding. In addition, all three OGG subfamilies bind 8-oxoG in a different manner even though the crucial interaction between the enzyme and the protonated N7 of 8-oxoG is conserved. Finally, the three OGG subfamilies differ with respect to DNA binding properties, helix-hairpin-helix motifs, and specificity for the opposite base. Full article
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology)
Open AccessReview The Role of Protein Crystallography in Defining the Mechanisms of Biogenesis and Catalysis in Copper Amine Oxidase
Int. J. Mol. Sci. 2012, 13(5), 5375-5405; doi:10.3390/ijms13055375
Received: 6 April 2012 / Revised: 22 April 2012 / Accepted: 26 April 2012 / Published: 3 May 2012
Cited by 10 | PDF Full-text (1105 KB) | HTML Full-text | XML Full-text
Abstract
Copper amine oxidases (CAOs) are a ubiquitous group of enzymes that catalyze the conversion of primary amines to aldehydes coupled to the reduction of O2 to H2O2. These enzymes utilize a wide range of substrates from methylamine to
[...] Read more.
Copper amine oxidases (CAOs) are a ubiquitous group of enzymes that catalyze the conversion of primary amines to aldehydes coupled to the reduction of O2 to H2O2. These enzymes utilize a wide range of substrates from methylamine to polypeptides. Changes in CAO activity are correlated with a variety of human diseases, including diabetes mellitus, Alzheimer’s disease, and inflammatory disorders. CAOs contain a cofactor, 2,4,5-trihydroxyphenylalanine quinone (TPQ), that is required for catalytic activity and synthesized through the post-translational modification of a tyrosine residue within the CAO polypeptide. TPQ generation is a self-processing event only requiring the addition of oxygen and Cu(II) to the apoCAO. Thus, the CAO active site supports two very different reactions: TPQ synthesis, and the two electron oxidation of primary amines. Crystal structures are available from bacterial through to human sources, and have given insight into substrate preference, stereospecificity, and structural changes during biogenesis and catalysis. In particular both these processes have been studied in crystallo through the addition of native substrates. These latter studies enable intermediates during physiological turnover to be directly visualized, and demonstrate the power of this relatively recent development in protein crystallography. Full article
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology)
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Open AccessReview Structural Features of Caspase-Activating Complexes
Int. J. Mol. Sci. 2012, 13(4), 4807-4818; doi:10.3390/ijms13044807
Received: 7 March 2012 / Revised: 28 March 2012 / Accepted: 10 April 2012 / Published: 16 April 2012
Cited by 24 | PDF Full-text (406 KB) | HTML Full-text | XML Full-text
Abstract
Apoptosis, also called programmed cell death, is an orderly cellular suicide program that is critical for the development, immune regulation and homeostasis of a multi-cellular organism. Failure to control this process can lead to serious human diseases, including many types of cancer, neurodegenerative
[...] Read more.
Apoptosis, also called programmed cell death, is an orderly cellular suicide program that is critical for the development, immune regulation and homeostasis of a multi-cellular organism. Failure to control this process can lead to serious human diseases, including many types of cancer, neurodegenerative diseases, and autoimmununity. The process of apoptosis is mediated by the sequential activation of caspases, which are cysteine proteases. Initiator caspases, such as caspase-2, -8, -9, and -10, are activated by formation of caspase-activating complexes, which function as a platform to recruit caspases, providing proximity for self-activation. Well-known initiator caspase-activating complexes include (1) DISC (Death Inducing Signaling Complex), which activates caspases-8 and 10; (2) Apoptosome, which activates caspase-9; and (3) PIDDosome, which activates caspase-2. Because of the fundamental biological importance of capases, many structural and biochemical studies to understand the molecular basis of assembly mechanism of caspase-activating complexes have been performed. In this review, we summarize previous studies that have examined the structural and biochemical features of caspase-activating complexes. By analyzing the structural basis for the assembly mechanism of the caspase-activating complex, we hope to provide a comprehensive understanding of caspase activation by these important oligomeric complexes. Full article
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology)

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