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Aspartic proteinase from barley grains is related to mammalian lysosomal cathepsin D

Sarkkinen, Paula; Kalkkinen, Nisse; Tilgmann, Carola LU ; Siuro, Jari; Kervinen, Jukka and Mikola, Leena (1992) In Planta 186(3). p.317-323
Abstract

Resting barley (Hordeum vulgare L.) grains contain acid-proteinase activity. The corresponding enzyme was purified from grain extracts by affinity chromatography on a pepstatin-Sepharose column. The pH optimum of the affinity-purified enzyme was between 3.5 and 3.9 as measured by hemoglobin hydrolysis and the enzymatic activity was completely inhibited by pepstatin a specific inhibitor of aspartic proteinases (EC 3.4.23). Further purification on a Mono S column followed by activity measurements and sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the affinity-purified enzyme preparation contained two active heterodimeric aspartic proteinases: a larger 48k Da enzyme, consisting of 32-kDa and 16-kDa subunits and a... (More)

Resting barley (Hordeum vulgare L.) grains contain acid-proteinase activity. The corresponding enzyme was purified from grain extracts by affinity chromatography on a pepstatin-Sepharose column. The pH optimum of the affinity-purified enzyme was between 3.5 and 3.9 as measured by hemoglobin hydrolysis and the enzymatic activity was completely inhibited by pepstatin a specific inhibitor of aspartic proteinases (EC 3.4.23). Further purification on a Mono S column followed by activity measurements and sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the affinity-purified enzyme preparation contained two active heterodimeric aspartic proteinases: a larger 48k Da enzyme, consisting of 32-kDa and 16-kDa subunits and a smaller one of 40 kDa, consisting of 29-kDa and 11-kDa subunits. Separation and partial amino acid sequence analysis of each subunit indicate that the 40-kDa enzyme is formed by proteolytic processing of the 48k Da form. Amino-acid sequence alignment and inhibition studies showed that the barley aspartic proteinase resembles mammalian lysosomal cathepsin D (EC 3.4.23.5). © 1992 Springer-Verlag.

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author
publishing date
type
Contribution to journal
publication status
published
keywords
Aspartic proteinase, Cathepsin D, Endopeptidase, Hordeum (proteinase)
in
Planta
volume
186
issue
3
pages
7 pages
publisher
Springer
external identifiers
  • Scopus:0000296581
ISSN
0032-0935
DOI
10.1007/BF00195311
language
English
LU publication?
no
id
0c25088a-eceb-4da9-8ce0-0d2b4f9dbf0f
date added to LUP
2016-04-11 13:24:51
date last changed
2016-11-27 04:39:17
@misc{0c25088a-eceb-4da9-8ce0-0d2b4f9dbf0f,
  abstract     = {<p>Resting barley (Hordeum vulgare L.) grains contain acid-proteinase activity. The corresponding enzyme was purified from grain extracts by affinity chromatography on a pepstatin-Sepharose column. The pH optimum of the affinity-purified enzyme was between 3.5 and 3.9 as measured by hemoglobin hydrolysis and the enzymatic activity was completely inhibited by pepstatin a specific inhibitor of aspartic proteinases (EC 3.4.23). Further purification on a Mono S column followed by activity measurements and sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the affinity-purified enzyme preparation contained two active heterodimeric aspartic proteinases: a larger 48k Da enzyme, consisting of 32-kDa and 16-kDa subunits and a smaller one of 40 kDa, consisting of 29-kDa and 11-kDa subunits. Separation and partial amino acid sequence analysis of each subunit indicate that the 40-kDa enzyme is formed by proteolytic processing of the 48k Da form. Amino-acid sequence alignment and inhibition studies showed that the barley aspartic proteinase resembles mammalian lysosomal cathepsin D (EC 3.4.23.5). © 1992 Springer-Verlag.</p>},
  author       = {Sarkkinen, Paula and Kalkkinen, Nisse and Tilgmann, Carola and Siuro, Jari and Kervinen, Jukka and Mikola, Leena},
  issn         = {0032-0935},
  keyword      = {Aspartic proteinase,Cathepsin D,Endopeptidase,Hordeum (proteinase)},
  language     = {eng},
  number       = {3},
  pages        = {317--323},
  publisher    = {ARRAY(0x8404330)},
  series       = {Planta},
  title        = {Aspartic proteinase from barley grains is related to mammalian lysosomal cathepsin D},
  url          = {http://dx.doi.org/10.1007/BF00195311},
  volume       = {186},
  year         = {1992},
}