The less conserved metal-binding site in human CRISP1 remains sensitive to zinc ions to permit protein oligomerization

Abstract Cysteine-rich secretory proteins (CRISPs) are a subgroup of the CRISP, antigen 5 and PR-1 (CAP) superfamily that is characterized by the presence of a conserved CAP domain. Two conserved histidines in the CAP domain are proposed to function as a Zn2+-binding site with unknown function. Huma...

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Autores principales:	Jie Sheng, Bart M. Gadella, Nick K. Olrichs, Dora V. Kaloyanova, J. Bernd Helms
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b17809b3b0aa4614bc98b150707df310
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spelling	oai:doaj.org-article:b17809b3b0aa4614bc98b150707df3102021-12-02T13:19:20ZThe less conserved metal-binding site in human CRISP1 remains sensitive to zinc ions to permit protein oligomerization10.1038/s41598-021-84926-y2045-2322https://doaj.org/article/b17809b3b0aa4614bc98b150707df3102021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-84926-yhttps://doaj.org/toc/2045-2322Abstract Cysteine-rich secretory proteins (CRISPs) are a subgroup of the CRISP, antigen 5 and PR-1 (CAP) superfamily that is characterized by the presence of a conserved CAP domain. Two conserved histidines in the CAP domain are proposed to function as a Zn2+-binding site with unknown function. Human CRISP1 is, however, one of the few family members that lack one of these characteristic histidine residues. The Zn2+-dependent oligomerization properties of human CRISP1 were investigated using a maltose-binding protein (MBP)-tagging approach in combination with low expression levels in XL-1 Blue bacteria. Moderate yields of soluble recombinant MBP-tagged human CRISP1 (MBP-CRISP1) and the MBP-tagged CAP domain of CRISP1 (MBP-CRISP1ΔC) were obtained. Zn2+ specifically induced oligomerization of both MBP-CRISP1 and MBP-CRISP1ΔC in vitro. The conserved His142 in the CAP domain was essential for this Zn2+ dependent oligomerization process, confirming a role of the CAP metal-binding site in the interaction with Zn2+. Furthermore, MBP-CRISP1 and MBP-CRISP1ΔC oligomers dissociated into monomers upon Zn2+ removal by EDTA. Condensation of proteins is characteristic for maturing sperm in the epididymis and this process was previously found to be Zn2+-dependent. The Zn2+-induced oligomerization of human recombinant CRISP1 may shed novel insights into the formation of functional protein complexes involved in mammalian fertilization.Jie ShengBart M. GadellaNick K. OlrichsDora V. KaloyanovaJ. Bernd HelmsNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jie Sheng Bart M. Gadella Nick K. Olrichs Dora V. Kaloyanova J. Bernd Helms The less conserved metal-binding site in human CRISP1 remains sensitive to zinc ions to permit protein oligomerization
description	Abstract Cysteine-rich secretory proteins (CRISPs) are a subgroup of the CRISP, antigen 5 and PR-1 (CAP) superfamily that is characterized by the presence of a conserved CAP domain. Two conserved histidines in the CAP domain are proposed to function as a Zn2+-binding site with unknown function. Human CRISP1 is, however, one of the few family members that lack one of these characteristic histidine residues. The Zn2+-dependent oligomerization properties of human CRISP1 were investigated using a maltose-binding protein (MBP)-tagging approach in combination with low expression levels in XL-1 Blue bacteria. Moderate yields of soluble recombinant MBP-tagged human CRISP1 (MBP-CRISP1) and the MBP-tagged CAP domain of CRISP1 (MBP-CRISP1ΔC) were obtained. Zn2+ specifically induced oligomerization of both MBP-CRISP1 and MBP-CRISP1ΔC in vitro. The conserved His142 in the CAP domain was essential for this Zn2+ dependent oligomerization process, confirming a role of the CAP metal-binding site in the interaction with Zn2+. Furthermore, MBP-CRISP1 and MBP-CRISP1ΔC oligomers dissociated into monomers upon Zn2+ removal by EDTA. Condensation of proteins is characteristic for maturing sperm in the epididymis and this process was previously found to be Zn2+-dependent. The Zn2+-induced oligomerization of human recombinant CRISP1 may shed novel insights into the formation of functional protein complexes involved in mammalian fertilization.
format	article
author	Jie Sheng Bart M. Gadella Nick K. Olrichs Dora V. Kaloyanova J. Bernd Helms
author_facet	Jie Sheng Bart M. Gadella Nick K. Olrichs Dora V. Kaloyanova J. Bernd Helms
author_sort	Jie Sheng
title	The less conserved metal-binding site in human CRISP1 remains sensitive to zinc ions to permit protein oligomerization
title_short	The less conserved metal-binding site in human CRISP1 remains sensitive to zinc ions to permit protein oligomerization
title_full	The less conserved metal-binding site in human CRISP1 remains sensitive to zinc ions to permit protein oligomerization
title_fullStr	The less conserved metal-binding site in human CRISP1 remains sensitive to zinc ions to permit protein oligomerization
title_full_unstemmed	The less conserved metal-binding site in human CRISP1 remains sensitive to zinc ions to permit protein oligomerization
title_sort	less conserved metal-binding site in human crisp1 remains sensitive to zinc ions to permit protein oligomerization
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/b17809b3b0aa4614bc98b150707df310
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The less conserved metal-binding site in human CRISP1 remains sensitive to zinc ions to permit protein oligomerization

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