Elucidation of the thermal stability of the neutral proteinase II from Aspergillus oryzae
| ISSN: | 0006-3002 |
|---|---|
| Přístupová URL adresa: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::170668857b183851c8ea1f3b14312688 https://pubmed.ncbi.nlm.nih.gov/8086433 |
| Rights: | CLOSED |
| Přírůstkové číslo: | edsair.doi.dedup.....170668857b183851c8ea1f3b14312688 |
| Autor: | Haruhide Kawabe, Hiroki Tatsumi, Hiroshi Motai, Eiichi Nakano, Kazuo Ikegaya, Seiji Murakami |
| Rok vydání: | 1994 |
| Předmět: |
Protein Folding
DNA Complementary Hot Temperature Stereochemistry Aspergillus oryzae Mutant Molecular Sequence Data Biophysics Gene Expression Saccharomyces cerevisiae Biochemistry Structure-Activity Relationship Structural Biology Thermolysin Enzyme Stability Escherichia coli Denaturation (biochemistry) Thermal stability Disulfides Site-directed mutagenesis Molecular Biology biology Base Sequence Chemistry Mutagenesis Gene Transfer Techniques Metalloendopeptidases biology.organism_classification Recombinant Proteins Mutagenesis Site-Directed Protein folding Electrophoresis Polyacrylamide Gel Plasmids |
| Zdroj: | Biochimica et biophysica acta. 1208(1) |
| ISSN: | 0006-3002 |
| Popis: | The neutral proteinase II from Aspergillus oryzae (NpII) is a zinc proteinase with three intramolecular disulfide bonds. NpII is most unstable after 10 min at about 75 degrees C, but regains stability beyond this temperature and is relatively stable at 100 degrees C. We analyzed the thermal stability of wild-type NpII and apo NpII. The results suggested that NpII unfolds reversibly upon incubation up to 100 degrees C, and that the irreversible inactivation observed is mainly due to autoproteolysis. To further understand the stability, a mutant NpII (Cys78-->Ala) lacking one of the disulfide bonds, was produced in a heterologous yeast expression system. The mutant NpII showed a similar stability profile, but the most unstable temperature and the most catalytically active temperature decreased to the same extent (around 10 degrees C), confirming that autoproteolysis is the main cause of the irreversible inactivation. Several lines of evidence presented in this study demonstrated that the thermal stability of o++NpII is attributed to reversible thermal unfolding and autoproteolysis. |
| Databáze: | OpenAIRE |
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