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Cited by 344Protein Structure and DynamicsProtein purification and stabilityEnzyme Structure and Function

Pro-sequence of subtilisin can guide the refolding of denatured subtilisin in an intermolecular process.

Zhu, Xueli, Ohta, Yoshiji, Jordan, Frank, Inouye, Masayori·1989·Nature
DOI 10.1038/339483a0
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Abstract

Subtilisin E, an alkaline serine protease consisting of a single polypeptide chain of 275 amino acids is produced from a pre-pro-protein. The pre-sequence functions as the signal peptide for protein secretion across the membrane. Deletion of the pro-sequence yields mature but inactive subtilisin: the 77-amino acid pro-sequence must precede the mature subtilisin to guide the latter into an active conformation. Pro-subtilisin denatured in 6 M guanidine-HCl can be self-processed to the active enzyme intramolecularly, with concomitant cleavage of the pro-sequence, when dialysed against renaturing buffer. We have constructed an active-centre mutant of pro-subtilisin (Asp 32----Asn) which is not processed to active enzyme, unlike the wild-type pro-subtilisin, because intramolecular processing is prevented. Here we report an intermolecular pathway for the refolding of the inactive mature protein to an active enzyme in vitro with the aid of exogenously added pro-sequence. We establish conditions under which the mature inactive form, as well as acid-denatured subtilisins Carlsberg and BPN', can be renatured by the mutant pro-subtilisin.

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Crossref · 10.1038/339483a0

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  • The pro-peptide of subtilisin is a catalyst for correct folding of the active enzyme.

    “The pro-peptide of subtilisin is a catalyst for correct folding of the active enzyme (Zhu et al., 1989).”

    From Modeled Structure of the Cell Envelope Proteinase of Lactococcus lactis (2020) · cites this paper as Zhu et al., 1989

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Authors
Zhu, X., Ohta, Y., Jordan, F., & Inouye, M.
Affiliations
Rutgers, The State University of New Jersey (US)
ORCID
Venue
Nature
Publisher
Springer Science and Business Media LLC
Published
1989-06
Volume
339
Issue
6224
Pages
483-484
ISSN
  • 0028-0836 (ISSN-L)
  • 1476-4687
DOI
10.1038/339483a0
OpenAlex
W2045786862
Cited by
344
References
10
Citations by year
2025: 2 · 2024: 2 · 2023: 4 · 2022: 3 · 2021: 2 · 2020: 4 · 2019: 2 · 2018: 3 · 2017: 3 · 2016: 3 · 2015: 5 · 2014: 8 · 2013: 4 · 2012: 10
FWCI
8.12
Type
journal article
Language
en
Open access
closed
Licence
www.springer.com/tdm (tdm)
Locations
  • Nature (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
  • Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
Keywords
subtilisin, protein refolding, protein denaturation, guanidine hydrochloride denaturation, serine protease activation, active site mutants, subtilisin Carlsberg, subtilisin BPN', renaturation
MeSH terms
Enzyme Activation, Enzyme Precursors, Escherichia coli, Genetic Vectors, Kinetics, Peptide Fragments, Plasmids, Protein Conformation, Protein Denaturation, Recombinant Proteins, Subtilisins

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