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Random substitution of large parts of the propeptide of yeast proteinase A

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Random substitution of large parts of the propeptide of yeast proteinase A. / van den Hazel, H B; Kielland-Brandt, Morten; Winther, Jakob R.

I: Journal of Biological Chemistry, Bind 270, Nr. 15, 1995, s. 8602-9.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Harvard

van den Hazel, HB, Kielland-Brandt, M & Winther, JR 1995, 'Random substitution of large parts of the propeptide of yeast proteinase A', Journal of Biological Chemistry, bind 270, nr. 15, s. 8602-9.

APA

van den Hazel, H. B., Kielland-Brandt, M., & Winther, J. R. (1995). Random substitution of large parts of the propeptide of yeast proteinase A. Journal of Biological Chemistry, 270(15), 8602-9.

Vancouver

van den Hazel HB, Kielland-Brandt M, Winther JR. Random substitution of large parts of the propeptide of yeast proteinase A. Journal of Biological Chemistry. 1995;270(15):8602-9.

Author

van den Hazel, H B ; Kielland-Brandt, Morten ; Winther, Jakob R. / Random substitution of large parts of the propeptide of yeast proteinase A. I: Journal of Biological Chemistry. 1995 ; Bind 270, Nr. 15. s. 8602-9.

Bibtex

@article{c59345cdf6ca47649409200b1262a5f1,
title = "Random substitution of large parts of the propeptide of yeast proteinase A",
abstract = "The yeast aspartic protease, proteinase A, has a 54 amino-acid propeptide, which is removed during activation of the zymogen in the vacuole. Apart from being involved inhibition/activation, the propeptide has been shown to be essential for formation of a stable active enzyme (van den Hazel, H. B., Kielland-Brandt, M. C., and Winther, J. R. (1993) J. Biol. Chem. 268, 18002-18007). We have investigated the sequence requirements for function of the propeptide. The N-terminal half and the C-terminal half of the propeptide were replaced by random sequences at the genetic level, and collections of the mutants were subjected to a colony screen for ones exhibiting activity. A high frequency (around 1{\%}) of active constructs was found, which indicates a very high tolerance for mutations in the propeptide. Thirty-nine functional mutant forms containing random sequence at either the N- or C-terminal half of the propeptide were characterized. Comparison of the propeptides of the active constructs suggests that a particular lysine residue is important for efficient biosynthesis of proteinase A.",
keywords = "Amino Acid Sequence, Aspartic Acid Endopeptidases, Base Sequence, Enzyme Precursors, Molecular Sequence Data, Mutation, Oligodeoxyribonucleotides, Peptide Fragments, Plasmids, Protein Precursors, Protein Processing, Post-Translational, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins",
author = "{van den Hazel}, {H B} and Morten Kielland-Brandt and Winther, {Jakob R.}",
year = "1995",
language = "English",
volume = "270",
pages = "8602--9",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology, Inc.",
number = "15",

}

RIS

TY - JOUR

T1 - Random substitution of large parts of the propeptide of yeast proteinase A

AU - van den Hazel, H B

AU - Kielland-Brandt, Morten

AU - Winther, Jakob R.

PY - 1995

Y1 - 1995

N2 - The yeast aspartic protease, proteinase A, has a 54 amino-acid propeptide, which is removed during activation of the zymogen in the vacuole. Apart from being involved inhibition/activation, the propeptide has been shown to be essential for formation of a stable active enzyme (van den Hazel, H. B., Kielland-Brandt, M. C., and Winther, J. R. (1993) J. Biol. Chem. 268, 18002-18007). We have investigated the sequence requirements for function of the propeptide. The N-terminal half and the C-terminal half of the propeptide were replaced by random sequences at the genetic level, and collections of the mutants were subjected to a colony screen for ones exhibiting activity. A high frequency (around 1%) of active constructs was found, which indicates a very high tolerance for mutations in the propeptide. Thirty-nine functional mutant forms containing random sequence at either the N- or C-terminal half of the propeptide were characterized. Comparison of the propeptides of the active constructs suggests that a particular lysine residue is important for efficient biosynthesis of proteinase A.

AB - The yeast aspartic protease, proteinase A, has a 54 amino-acid propeptide, which is removed during activation of the zymogen in the vacuole. Apart from being involved inhibition/activation, the propeptide has been shown to be essential for formation of a stable active enzyme (van den Hazel, H. B., Kielland-Brandt, M. C., and Winther, J. R. (1993) J. Biol. Chem. 268, 18002-18007). We have investigated the sequence requirements for function of the propeptide. The N-terminal half and the C-terminal half of the propeptide were replaced by random sequences at the genetic level, and collections of the mutants were subjected to a colony screen for ones exhibiting activity. A high frequency (around 1%) of active constructs was found, which indicates a very high tolerance for mutations in the propeptide. Thirty-nine functional mutant forms containing random sequence at either the N- or C-terminal half of the propeptide were characterized. Comparison of the propeptides of the active constructs suggests that a particular lysine residue is important for efficient biosynthesis of proteinase A.

KW - Amino Acid Sequence

KW - Aspartic Acid Endopeptidases

KW - Base Sequence

KW - Enzyme Precursors

KW - Molecular Sequence Data

KW - Mutation

KW - Oligodeoxyribonucleotides

KW - Peptide Fragments

KW - Plasmids

KW - Protein Precursors

KW - Protein Processing, Post-Translational

KW - Saccharomyces cerevisiae

KW - Saccharomyces cerevisiae Proteins

M3 - Journal article

C2 - 7721762

VL - 270

SP - 8602

EP - 8609

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 15

ER -

ID: 43974368