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Complexome Profiling: Assembly and Remodeling of Protein Complexes

Butyrylcholinesterase–Protein Interactions in Human Serum

Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Medical University of Gdańsk, 80-416 Gdańsk, Poland
Department of Pharmaceutical Pathophysiology, Faculty of Pharmacy, Medical University of Gdańsk, 80-416 Gdańsk, Poland
Mass Spectrometry Laboratory Institute of Biochemistry and Biophysics, Polish Academy of Sciences, 02-106 Warsaw, Poland
Department of Laboratory Medicine, Medical University of Gdańsk, 80-952 Gdańsk, Poland
Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdańsk, Wita Stwosza 63, 80-308 Gdańsk, Poland
Department of Biology and Medical Genetics, Medical University of Gdańsk, 80-210 Gdańsk, Poland
Laboratory of Clinical Genetics, University Clinical Centre, 80-952 Gdańsk, Poland
Author to whom correspondence should be addressed.
Academic Editor: Istvan Simon
Int. J. Mol. Sci. 2021, 22(19), 10662;
Received: 12 August 2021 / Revised: 24 September 2021 / Accepted: 28 September 2021 / Published: 1 October 2021
Measuring various biochemical and cellular components in the blood is a routine procedure in clinical practice. Human serum contains hundreds of diverse proteins secreted from all cells and tissues in healthy and diseased states. Moreover, some serum proteins have specific strong interactions with other blood components, but most interactions are probably weak and transient. One of the serum proteins is butyrylcholinesterase (BChE), an enzyme existing mainly as a glycosylated soluble tetramer that plays an important role in the metabolism of many drugs. Our results suggest that BChE interacts with plasma proteins and forms much larger complexes than predicted from the molecular weight of the BChE tetramer. To investigate and isolate such complexes, we developed a two-step strategy to find specific protein–protein interactions by combining native size-exclusion chromatography (SEC) with affinity chromatography with the resin that specifically binds BChE. Second, to confirm protein complexes′ specificity, we fractionated blood serum proteins by density gradient ultracentrifugation followed by co-immunoprecipitation with anti-BChE monoclonal antibodies. The proteins coisolated in complexes with BChE were identified by mass spectroscopy. These binding studies revealed that BChE interacts with a number of proteins in the human serum. Some of these interactions seem to be more stable than transient. BChE copurification with ApoA-I and the density of some fractions containing BChE corresponding to high-density lipoprotein cholesterol (HDL) during ultracentrifugation suggest its interactions with HDL. Moreover, we observed lower BChE plasma activity in individuals with severely reduced HDL levels (≤20 mg/dL). The presented two-step methodology for determination of the BChE interactions can facilitate further analysis of such complexes, especially from the brain tissue, where BChE could be involved in the pathogenesis and progression of AD. View Full-Text
Keywords: BChE; butyrylcholinesterase; pseudocholinesterase; high-density lipoprotein (HDL); protein interactions; ApoA-I; protein interactions BChE; butyrylcholinesterase; pseudocholinesterase; high-density lipoprotein (HDL); protein interactions; ApoA-I; protein interactions
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MDPI and ACS Style

Jasiecki, J.; Szczoczarz, A.; Cysewski, D.; Lewandowski, K.; Skowron, P.; Waleron, K.; Wasąg, B. Butyrylcholinesterase–Protein Interactions in Human Serum. Int. J. Mol. Sci. 2021, 22, 10662.

AMA Style

Jasiecki J, Szczoczarz A, Cysewski D, Lewandowski K, Skowron P, Waleron K, Wasąg B. Butyrylcholinesterase–Protein Interactions in Human Serum. International Journal of Molecular Sciences. 2021; 22(19):10662.

Chicago/Turabian Style

Jasiecki, Jacek, Anna Szczoczarz, Dominik Cysewski, Krzysztof Lewandowski, Piotr Skowron, Krzysztof Waleron, and Bartosz Wasąg. 2021. "Butyrylcholinesterase–Protein Interactions in Human Serum" International Journal of Molecular Sciences 22, no. 19: 10662.

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