TY - JOUR

T1 - The propeptide is required for in vivo formation of stable active yeast proteinase A and can function even when not covalently linked to the mature region

AU - van den Hazel, H B

AU - Kielland-Brandt, Morten

AU - Winther, Jakob R.

PY - 1993

Y1 - 1993

N2 - The PEP4-encoded aspartate protease proteinase A from Saccharomyces cerevisiae is synthesized as a zymogen (Ammerer, G., Hunter, C. P., Rothman, J. H., Saari, G. C., Valls, L. A., and Stevens, T. H. (1986) Mol. Cell. Biol. 6, 2490-2499; Woolford, C. A., Daniels, L. B., Park, F. J., Jones, E. W., van Arsdell, J. N., and Innis, M. A. (1986) Mol. Cell. Biol. 6, 2500-2510). We constructed a mutant form, lacking the propeptide. This form, still containing the signal peptide, was translocated into the endoplasmic reticulum, but the mature region was subsequently completely degraded. When a plasmid encoding only the propeptide after the signal peptide was introduced into a strain producing the mature region, a subpopulation of mature region molecules was rescued from the degradation and gained activity. Increased activity was found when the mature region was co-produced with increased amounts of propeptide, whereas truncated propeptides, lacking residues at its C terminus, were less efficient in the interaction with the mature region. We propose that the mature region of proteinase A cannot fold into its stable active conformation in the absence of the propeptide and that the propeptide can promote folding of the mature region, even when the propeptide and the mature region are not covalently linked.

AB - The PEP4-encoded aspartate protease proteinase A from Saccharomyces cerevisiae is synthesized as a zymogen (Ammerer, G., Hunter, C. P., Rothman, J. H., Saari, G. C., Valls, L. A., and Stevens, T. H. (1986) Mol. Cell. Biol. 6, 2490-2499; Woolford, C. A., Daniels, L. B., Park, F. J., Jones, E. W., van Arsdell, J. N., and Innis, M. A. (1986) Mol. Cell. Biol. 6, 2500-2510). We constructed a mutant form, lacking the propeptide. This form, still containing the signal peptide, was translocated into the endoplasmic reticulum, but the mature region was subsequently completely degraded. When a plasmid encoding only the propeptide after the signal peptide was introduced into a strain producing the mature region, a subpopulation of mature region molecules was rescued from the degradation and gained activity. Increased activity was found when the mature region was co-produced with increased amounts of propeptide, whereas truncated propeptides, lacking residues at its C terminus, were less efficient in the interaction with the mature region. We propose that the mature region of proteinase A cannot fold into its stable active conformation in the absence of the propeptide and that the propeptide can promote folding of the mature region, even when the propeptide and the mature region are not covalently linked.

KW - Alleles

KW - Amino Acid Sequence

KW - Aspartic Acid Endopeptidases

KW - Base Sequence

KW - Cloning, Molecular

KW - Enzyme Precursors

KW - Fungal Proteins

KW - Genes, Fungal

KW - Molecular Sequence Data

KW - Mutagenesis

KW - Oligodeoxyribonucleotides

KW - Plasmids

KW - Recombinant Proteins

KW - Restriction Mapping

KW - Saccharomyces cerevisiae

KW - Saccharomyces cerevisiae Proteins

M3 - Journal article

C2 - 8349680

VL - 268

SP - 18002

EP - 18007

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 24

ER -