Cargando…

Structural and biochemical characterisation of Co(2+)-binding sites on serum albumins and their interplay with fatty acids

Serum albumin–Co(2+) interactions are of clinical importance. They play a role in mediating the physiological effects associated with cobalt toxicity and are central to the albumin cobalt binding (ACB) assay for diagnosis of myocardial ischemia. To further understand these processes, a deeper unders...

Descripción completa

Detalles Bibliográficos
Autores principales: Wu, Dongmei, Gucwa, Michal, Czub, Mateusz P., Cooper, David R., Shabalin, Ivan G., Fritzen, Remi, Arya, Swati, Schwarz-Linek, Ulrich, Blindauer, Claudia A., Minor, Wladek, Stewart, Alan J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society of Chemistry 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10266443/
https://www.ncbi.nlm.nih.gov/pubmed/37325156
http://dx.doi.org/10.1039/d3sc01723k
_version_ 1785058741676146688
author Wu, Dongmei
Gucwa, Michal
Czub, Mateusz P.
Cooper, David R.
Shabalin, Ivan G.
Fritzen, Remi
Arya, Swati
Schwarz-Linek, Ulrich
Blindauer, Claudia A.
Minor, Wladek
Stewart, Alan J.
author_facet Wu, Dongmei
Gucwa, Michal
Czub, Mateusz P.
Cooper, David R.
Shabalin, Ivan G.
Fritzen, Remi
Arya, Swati
Schwarz-Linek, Ulrich
Blindauer, Claudia A.
Minor, Wladek
Stewart, Alan J.
author_sort Wu, Dongmei
collection PubMed
description Serum albumin–Co(2+) interactions are of clinical importance. They play a role in mediating the physiological effects associated with cobalt toxicity and are central to the albumin cobalt binding (ACB) assay for diagnosis of myocardial ischemia. To further understand these processes, a deeper understanding of albumin–Co(2+) interactions is required. Here, we present the first crystallographic structures of human serum albumin (HSA; three structures) and equine serum albumin (ESA; one structure) in complex with Co(2+). Amongst a total of sixteen sites bearing a cobalt ion across the structures, two locations were prominent, and they relate to metal-binding sites A and B. Site-directed mutagenesis and isothermal titration calorimetry (ITC) were employed to characterise sites on HSA. The results indicate that His9 and His67 contribute to the primary (putatively corresponding to site B) and secondary Co(2+)-binding sites (site A), respectively. The presence of additional multiple weak-affinity Co(2+) binding sites on HSA was also supported by ITC studies. Furthermore, addition of 5 molar equivalents of the non-esterified fatty acid palmitate (C16:0) reduced the Co(2+)-binding affinity at both sites A and B. The presence of bound myristate (C14:0) in the HSA crystal structures provided insight into the fatty acid-mediated structural changes that diminish the affinity of the protein toward Co(2+). Together, these data provide further support for the idea that ischemia-modified albumin corresponds to albumin with excessive fatty-acid loading. Collectively, our findings provide a comprehensive understanding of the molecular underpinnings governing Co(2+) binding to serum albumin.
format Online
Article
Text
id pubmed-10266443
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher The Royal Society of Chemistry
record_format MEDLINE/PubMed
spelling pubmed-102664432023-06-15 Structural and biochemical characterisation of Co(2+)-binding sites on serum albumins and their interplay with fatty acids Wu, Dongmei Gucwa, Michal Czub, Mateusz P. Cooper, David R. Shabalin, Ivan G. Fritzen, Remi Arya, Swati Schwarz-Linek, Ulrich Blindauer, Claudia A. Minor, Wladek Stewart, Alan J. Chem Sci Chemistry Serum albumin–Co(2+) interactions are of clinical importance. They play a role in mediating the physiological effects associated with cobalt toxicity and are central to the albumin cobalt binding (ACB) assay for diagnosis of myocardial ischemia. To further understand these processes, a deeper understanding of albumin–Co(2+) interactions is required. Here, we present the first crystallographic structures of human serum albumin (HSA; three structures) and equine serum albumin (ESA; one structure) in complex with Co(2+). Amongst a total of sixteen sites bearing a cobalt ion across the structures, two locations were prominent, and they relate to metal-binding sites A and B. Site-directed mutagenesis and isothermal titration calorimetry (ITC) were employed to characterise sites on HSA. The results indicate that His9 and His67 contribute to the primary (putatively corresponding to site B) and secondary Co(2+)-binding sites (site A), respectively. The presence of additional multiple weak-affinity Co(2+) binding sites on HSA was also supported by ITC studies. Furthermore, addition of 5 molar equivalents of the non-esterified fatty acid palmitate (C16:0) reduced the Co(2+)-binding affinity at both sites A and B. The presence of bound myristate (C14:0) in the HSA crystal structures provided insight into the fatty acid-mediated structural changes that diminish the affinity of the protein toward Co(2+). Together, these data provide further support for the idea that ischemia-modified albumin corresponds to albumin with excessive fatty-acid loading. Collectively, our findings provide a comprehensive understanding of the molecular underpinnings governing Co(2+) binding to serum albumin. The Royal Society of Chemistry 2023-05-23 /pmc/articles/PMC10266443/ /pubmed/37325156 http://dx.doi.org/10.1039/d3sc01723k Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by/3.0/
spellingShingle Chemistry
Wu, Dongmei
Gucwa, Michal
Czub, Mateusz P.
Cooper, David R.
Shabalin, Ivan G.
Fritzen, Remi
Arya, Swati
Schwarz-Linek, Ulrich
Blindauer, Claudia A.
Minor, Wladek
Stewart, Alan J.
Structural and biochemical characterisation of Co(2+)-binding sites on serum albumins and their interplay with fatty acids
title Structural and biochemical characterisation of Co(2+)-binding sites on serum albumins and their interplay with fatty acids
title_full Structural and biochemical characterisation of Co(2+)-binding sites on serum albumins and their interplay with fatty acids
title_fullStr Structural and biochemical characterisation of Co(2+)-binding sites on serum albumins and their interplay with fatty acids
title_full_unstemmed Structural and biochemical characterisation of Co(2+)-binding sites on serum albumins and their interplay with fatty acids
title_short Structural and biochemical characterisation of Co(2+)-binding sites on serum albumins and their interplay with fatty acids
title_sort structural and biochemical characterisation of co(2+)-binding sites on serum albumins and their interplay with fatty acids
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10266443/
https://www.ncbi.nlm.nih.gov/pubmed/37325156
http://dx.doi.org/10.1039/d3sc01723k
work_keys_str_mv AT wudongmei structuralandbiochemicalcharacterisationofco2bindingsitesonserumalbuminsandtheirinterplaywithfattyacids
AT gucwamichal structuralandbiochemicalcharacterisationofco2bindingsitesonserumalbuminsandtheirinterplaywithfattyacids
AT czubmateuszp structuralandbiochemicalcharacterisationofco2bindingsitesonserumalbuminsandtheirinterplaywithfattyacids
AT cooperdavidr structuralandbiochemicalcharacterisationofco2bindingsitesonserumalbuminsandtheirinterplaywithfattyacids
AT shabalinivang structuralandbiochemicalcharacterisationofco2bindingsitesonserumalbuminsandtheirinterplaywithfattyacids
AT fritzenremi structuralandbiochemicalcharacterisationofco2bindingsitesonserumalbuminsandtheirinterplaywithfattyacids
AT aryaswati structuralandbiochemicalcharacterisationofco2bindingsitesonserumalbuminsandtheirinterplaywithfattyacids
AT schwarzlinekulrich structuralandbiochemicalcharacterisationofco2bindingsitesonserumalbuminsandtheirinterplaywithfattyacids
AT blindauerclaudiaa structuralandbiochemicalcharacterisationofco2bindingsitesonserumalbuminsandtheirinterplaywithfattyacids
AT minorwladek structuralandbiochemicalcharacterisationofco2bindingsitesonserumalbuminsandtheirinterplaywithfattyacids
AT stewartalanj structuralandbiochemicalcharacterisationofco2bindingsitesonserumalbuminsandtheirinterplaywithfattyacids