Autoactivation of proteinase A initiates activation of yeast vacuolar zymogens

H B van den Hazel; Morten Kielland-Brandt; Jakob R. Winther

Autoactivation of proteinase A initiates activation of yeast vacuolar zymogens

H B van den Hazel, Morten Kielland-Brandt, Jakob R. Winther

Biomolecular Sciences

56 Citationer (Scopus)

Abstract

The Saccharomyces cerevisiae PEP4 gene encodes proteinase A, an aspartyl protease. pep4 mutants are defective in the activation of many vacuolar hydrolases, including proteinase B. We have expressed a pep4 mutation which directs the accumulation of pro-proteinase A with a defective active site. Co-expression with PEP4 leads to normal processing, i.e. the mutant zymogen is functional as a substrate for the maturation reaction in trans. We conclude that wild-type pro-proteinase A has the ability to mediate its own activation. Elimination of the co-expressed PEP4 gene did not effectively stop the processing of the mutant zymogen, owing to a strong, proteinase-B-dependent, phenotypic lag. In a proteinase-B-negative strain, processing of pro-proteinase A led to an active form of a higher molecular mass than the normal mature form.

Originalsprog	Engelsk
Tidsskrift	European Journal of Biochemistry
Vol/bind	207
Udgave nummer	1
Sider (fra-til)	277-83
Antal sider	7
ISSN	0014-2956
Status	Udgivet - 1992

Citationsformater

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title = "Autoactivation of proteinase A initiates activation of yeast vacuolar zymogens",

abstract = "The Saccharomyces cerevisiae PEP4 gene encodes proteinase A, an aspartyl protease. pep4 mutants are defective in the activation of many vacuolar hydrolases, including proteinase B. We have expressed a pep4 mutation which directs the accumulation of pro-proteinase A with a defective active site. Co-expression with PEP4 leads to normal processing, i.e. the mutant zymogen is functional as a substrate for the maturation reaction in trans. We conclude that wild-type pro-proteinase A has the ability to mediate its own activation. Elimination of the co-expressed PEP4 gene did not effectively stop the processing of the mutant zymogen, owing to a strong, proteinase-B-dependent, phenotypic lag. In a proteinase-B-negative strain, processing of pro-proteinase A led to an active form of a higher molecular mass than the normal mature form.",

keywords = "Alleles, Amino Acid Sequence, Aspartic Acid Endopeptidases, Base Sequence, Binding Sites, Codon, Enzyme Activation, Enzyme Precursors, Genes, Fungal, Molecular Sequence Data, Mutagenesis, Site-Directed, Phenotype, Plasmids, Polymerase Chain Reaction, Protein Processing, Post-Translational, Restriction Mapping, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Vacuoles",

author = "{van den Hazel}, {H B} and Morten Kielland-Brandt and Winther, {Jakob R.}",

year = "1992",

language = "English",

volume = "207",

pages = "277--83",

journal = "European Journal of Biochemistry",

issn = "0014-2956",

publisher = "Springer Verlag",

number = "1",

}

TY - JOUR

T1 - Autoactivation of proteinase A initiates activation of yeast vacuolar zymogens

AU - van den Hazel, H B

AU - Kielland-Brandt, Morten

AU - Winther, Jakob R.

PY - 1992

Y1 - 1992

N2 - The Saccharomyces cerevisiae PEP4 gene encodes proteinase A, an aspartyl protease. pep4 mutants are defective in the activation of many vacuolar hydrolases, including proteinase B. We have expressed a pep4 mutation which directs the accumulation of pro-proteinase A with a defective active site. Co-expression with PEP4 leads to normal processing, i.e. the mutant zymogen is functional as a substrate for the maturation reaction in trans. We conclude that wild-type pro-proteinase A has the ability to mediate its own activation. Elimination of the co-expressed PEP4 gene did not effectively stop the processing of the mutant zymogen, owing to a strong, proteinase-B-dependent, phenotypic lag. In a proteinase-B-negative strain, processing of pro-proteinase A led to an active form of a higher molecular mass than the normal mature form.

AB - The Saccharomyces cerevisiae PEP4 gene encodes proteinase A, an aspartyl protease. pep4 mutants are defective in the activation of many vacuolar hydrolases, including proteinase B. We have expressed a pep4 mutation which directs the accumulation of pro-proteinase A with a defective active site. Co-expression with PEP4 leads to normal processing, i.e. the mutant zymogen is functional as a substrate for the maturation reaction in trans. We conclude that wild-type pro-proteinase A has the ability to mediate its own activation. Elimination of the co-expressed PEP4 gene did not effectively stop the processing of the mutant zymogen, owing to a strong, proteinase-B-dependent, phenotypic lag. In a proteinase-B-negative strain, processing of pro-proteinase A led to an active form of a higher molecular mass than the normal mature form.

KW - Alleles

KW - Amino Acid Sequence

KW - Aspartic Acid Endopeptidases

KW - Base Sequence

KW - Binding Sites

KW - Codon

KW - Enzyme Activation

KW - Enzyme Precursors

KW - Genes, Fungal

KW - Molecular Sequence Data

KW - Mutagenesis, Site-Directed

KW - Phenotype

KW - Plasmids

KW - Polymerase Chain Reaction

KW - Protein Processing, Post-Translational

KW - Restriction Mapping

KW - Saccharomyces cerevisiae

KW - Saccharomyces cerevisiae Proteins

KW - Vacuoles

M3 - Journal article

C2 - 1628653

SN - 0014-2956

VL - 207

SP - 277

EP - 283

JO - European Journal of Biochemistry

JF - European Journal of Biochemistry

IS - 1

ER -

Autoactivation of proteinase A initiates activation of yeast vacuolar zymogens

Abstract

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