pH-dependent processing of yeast procarboxypeptidase Y by proteinase A in vivo and in vitro
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pH-dependent processing of yeast procarboxypeptidase Y by proteinase A in vivo and in vitro. / Sørensen, S O; van den Hazel, H B; Kielland-Brandt, Morten; Winther, Jakob R.
In: European Journal of Biochemistry, Vol. 220, No. 1, 1994, p. 19-27.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - pH-dependent processing of yeast procarboxypeptidase Y by proteinase A in vivo and in vitro
AU - Sørensen, S O
AU - van den Hazel, H B
AU - Kielland-Brandt, Morten
AU - Winther, Jakob R.
PY - 1994
Y1 - 1994
N2 - Carboxypeptidase Y is a vacuolar enzyme from Saccharomyces cerevisiae. It enters the vacuole as a zymogen, procarboxypeptidase Y, which is immediately processed in a reaction involving two endoproteases, proteinase A and proteinase B. We have investigated the in vitro activation of purified procarboxypeptidase Y by purified proteinase A. This has identified two different processing intermediates; one active and one inactive. The intermediates define a 33 amino acid segment of the 91 amino acid propeptide as sufficient for maintaining the enzyme in an inactive state. The inactive intermediate was isolated from a processing reaction at neutral pH. In order to investigate the influence of vacuolar pH on processing in vivo, the autoactivation of proteinase A and its processing of procarboxypeptidase Y were studied in a vma2 prb1 mutant, which is deficient in vacuolar acidification and proteinase B activity. Efficient processing of procarboxypeptidase Y in the absence of proteinase B is dependent on acidic vacuolar pH, and the processing at neutral pH is slow and takes place in two steps similar to those identified in vitro.
AB - Carboxypeptidase Y is a vacuolar enzyme from Saccharomyces cerevisiae. It enters the vacuole as a zymogen, procarboxypeptidase Y, which is immediately processed in a reaction involving two endoproteases, proteinase A and proteinase B. We have investigated the in vitro activation of purified procarboxypeptidase Y by purified proteinase A. This has identified two different processing intermediates; one active and one inactive. The intermediates define a 33 amino acid segment of the 91 amino acid propeptide as sufficient for maintaining the enzyme in an inactive state. The inactive intermediate was isolated from a processing reaction at neutral pH. In order to investigate the influence of vacuolar pH on processing in vivo, the autoactivation of proteinase A and its processing of procarboxypeptidase Y were studied in a vma2 prb1 mutant, which is deficient in vacuolar acidification and proteinase B activity. Efficient processing of procarboxypeptidase Y in the absence of proteinase B is dependent on acidic vacuolar pH, and the processing at neutral pH is slow and takes place in two steps similar to those identified in vitro.
KW - Amino Acid Sequence
KW - Aspartic Acid Endopeptidases
KW - Binding Sites
KW - Carboxypeptidases
KW - Cathepsin A
KW - Enzyme Activation
KW - Enzyme Precursors
KW - Genes, Fungal
KW - Hydrogen-Ion Concentration
KW - Molecular Sequence Data
KW - Mutation
KW - Protein Processing, Post-Translational
KW - Saccharomyces cerevisiae
KW - Saccharomyces cerevisiae Proteins
KW - Serine Endopeptidases
KW - Vacuoles
M3 - Journal article
C2 - 8119286
VL - 220
SP - 19
EP - 27
JO - FEBS Journal
JF - FEBS Journal
SN - 1742-464X
IS - 1
ER -
ID: 43974456