Next Article in Journal
Effects of Melatonin and Its Analogues on Pancreatic Inflammation, Enzyme Secretion, and Tumorigenesis
Next Article in Special Issue
Deciphering Structural Photophysics of Fluorescent Proteins by Kinetic Crystallography
Previous Article in Journal
Integrative RNA- and miRNA-Profile Analysis Reveals a Likely Role of BR and Auxin Signaling in Branch Angle Regulation of B. napus
Article Menu
Issue 5 (May) cover image

Export Article

Open AccessArticle
Int. J. Mol. Sci. 2017, 18(5), 1009; doi:10.3390/ijms18051009

Interaction of Biliverdin Chromophore with Near-Infrared Fluorescent Protein BphP1-FP Engineered from Bacterial Phytochrome

1
Laboratory of Structural Dynamics, Stability and Folding of Proteins, Institute of Cytology, Russian Academy of Sciences, 4 Tikhoretsky ave., St. Petersburg 194064, Russian
2
Department of Biophysics, Peter the Great St. Petersburg Polytechnic University, 29 Polytechnicheskaya st., St. Petersburg 195251, Russian
3
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, 1300 Morris Park ave., Bronx, NY 10461, USA
4
Department of Biochemistry and Developmental Biology, Faculty of Medicine, University of Helsinki, 8 Haartmaninkatu st., Helsinki 00290, Finland
*
Authors to whom correspondence should be addressed.
Academic Editors: Dominique Bourgeois and Hideaki Mizuno
Received: 12 April 2017 / Revised: 30 April 2017 / Accepted: 4 May 2017 / Published: 8 May 2017
(This article belongs to the Special Issue Fluorescent Proteins)
View Full-Text   |   Download PDF [4765 KB, uploaded 8 May 2017]   |  

Abstract

Near-infrared (NIR) fluorescent proteins (FPs) designed from PAS (Per-ARNT-Sim repeats) and GAF (cGMP phosphodiesterase/adenylate cyclase/FhlA transcriptional activator) domains of bacterial phytochromes covalently bind biliverdin (BV) chromophore via one or two Cys residues. We studied BV interaction with a series of NIR FP variants derived from the recently reported BphP1-FP protein. The latter was engineered from a bacterial phytochrome RpBphP1, and has two reactive Cys residues (Cys15 in the PAS domain and Cys256 in the GAF domain), whereas its mutants contain single Cys residues either in the PAS domain or in the GAF domain, or no Cys residues. We characterized BphP1-FP and its mutants biochemically and spectroscopically in the absence and in the presence of denaturant. We found that all BphP1-FP variants are monomers. We revealed that spectral properties of the BphP1-FP variants containing either Cys15 or Cys256, or both, are determined by the covalently bound BV chromophore only. Consequently, this suggests an involvement of the inter-monomeric allosteric effects in the BV interaction with monomers in dimeric NIR FPs, such as iRFPs. Likely, insertion of the Cys15 residue, in addition to the Cys256 residue, in dimeric NIR FPs influences BV binding by promoting the BV chromophore covalent cross-linking to both PAS and GAF domains. View Full-Text
Keywords: bacterial phytochrome; iRFP; IFP; near-infrared fluorescent protein; GAF domain; biliverdin; competitive inhibition bacterial phytochrome; iRFP; IFP; near-infrared fluorescent protein; GAF domain; biliverdin; competitive inhibition
Figures

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

MDPI and ACS Style

Stepanenko, O.V.; Stepanenko, O.V.; Kuznetsova, I.M.; Shcherbakova, D.M.; Verkhusha, V.V.; Turoverov, K.K. Interaction of Biliverdin Chromophore with Near-Infrared Fluorescent Protein BphP1-FP Engineered from Bacterial Phytochrome. Int. J. Mol. Sci. 2017, 18, 1009.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top