The less conserved metal-binding site in human CRISP1 remains sensitive to zinc ions to permit protein oligomerization
| Autor: | Sheng, Jie, Gadella, Bart M, Olrichs, Nick K, Kaloyanova, Dora V, Helms, J Bernd, LS Veterinaire biochemie, FAH klinische reproductie, Veterinaire biochemie, dB&C FR-RMSC FR, dES/dFAH FR, dB&C FR-RMSC RMSC |
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| Přispěvatelé: | LS Veterinaire biochemie, FAH klinische reproductie, Veterinaire biochemie, dB&C FR-RMSC FR, dES/dFAH FR, dB&C FR-RMSC RMSC |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
inorganic chemicals
Science Metal Binding Site Protein aggregation Biochemistry Article law.invention law Protein oligomerization Humans General Histidine Multidisciplinary Binding Sites Membrane Glycoproteins Chemistry In vitro Recombinant Proteins Zinc Secretory protein Recombinant DNA Medicine Protein Multimerization Function (biology) |
| Zdroj: | Scientific Reports, 11(1). NLM (Medline) Scientific Reports, Vol 11, Iss 1, Pp 1-10 (2021) Scientific Reports |
| ISSN: | 2045-2322 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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