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Butyrylcholinesterase–Protein Interactions in Human Serum
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 o...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8508650/ https://www.ncbi.nlm.nih.gov/pubmed/34639003 http://dx.doi.org/10.3390/ijms221910662 |
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author | Jasiecki, Jacek Szczoczarz, Anna Cysewski, Dominik Lewandowski, Krzysztof Skowron, Piotr Waleron, Krzysztof Wasąg, Bartosz |
author_facet | Jasiecki, Jacek Szczoczarz, Anna Cysewski, Dominik Lewandowski, Krzysztof Skowron, Piotr Waleron, Krzysztof Wasąg, Bartosz |
author_sort | Jasiecki, Jacek |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-8508650 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-85086502021-10-13 Butyrylcholinesterase–Protein Interactions in Human Serum Jasiecki, Jacek Szczoczarz, Anna Cysewski, Dominik Lewandowski, Krzysztof Skowron, Piotr Waleron, Krzysztof Wasąg, Bartosz Int J Mol Sci Article 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. MDPI 2021-10-01 /pmc/articles/PMC8508650/ /pubmed/34639003 http://dx.doi.org/10.3390/ijms221910662 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Jasiecki, Jacek Szczoczarz, Anna Cysewski, Dominik Lewandowski, Krzysztof Skowron, Piotr Waleron, Krzysztof Wasąg, Bartosz Butyrylcholinesterase–Protein Interactions in Human Serum |
title | Butyrylcholinesterase–Protein Interactions in Human Serum |
title_full | Butyrylcholinesterase–Protein Interactions in Human Serum |
title_fullStr | Butyrylcholinesterase–Protein Interactions in Human Serum |
title_full_unstemmed | Butyrylcholinesterase–Protein Interactions in Human Serum |
title_short | Butyrylcholinesterase–Protein Interactions in Human Serum |
title_sort | butyrylcholinesterase–protein interactions in human serum |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8508650/ https://www.ncbi.nlm.nih.gov/pubmed/34639003 http://dx.doi.org/10.3390/ijms221910662 |
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