Identification of the role of a cysteine-rich region of PC6B by determining the enzymatic characteristics of its mutants
| Autor: | Guan-Zhen Yang, Xiang-Fu Wu, Lie Wang |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Baculoviridae
Mutant chemistry.chemical_element Bioengineering Biology Calcium medicine.disease_cause Protein Engineering Applied Microbiology and Biotechnology Biochemistry Mice Enzyme Stability medicine Animals Cysteine Molecular Biology Gene Cells Cultured chemistry.chemical_classification Mutation Hydrogen-Ion Concentration biology.organism_classification Protein Structure Tertiary Kinetics Enzyme chemistry Proprotein Convertase 5 Proprotein Convertases Biotechnology |
| Zdroj: | Molecular biotechnology. 27(1) |
| ISSN: | 1073-6085 |
| Popis: | In the proprotein convertases family, mouse PC6B (mPC6B) has a very large cysteine-rich region (CRR), consisting of 22 tandem cysteine-rich (Cys-rich) repeated segments. The role of this region remains elusive. In this report, to get insight on the possible role of the CRR, we constructed four truncated mPC6B mutant genes with 0, 5, 11, and 22 Cys-rich repeated segments remaining; using the baculovirus-expression system and a simple purification method, we obtained four enzyme mutants of mPC6B. By determining their optimal pH and calcium ion concentration for enzymatic activity and their thermal stability, we found that CRR did not affect pH optimum and Ca2+ optimum compared with the p-domain. However, CRR acted as a stabilizing domain in addition to the p-domain. By kinetic analyses of four mutants, we found that the long Cys-rich repeats in the native form of mPC6B reduced its Vmax. These facts suggest that CRR acts as an important part of functional domain. |
| Databáze: | OpenAIRE |
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