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).
(Less)
- author
- Sarkkinen, Paula
; Kalkkinen, Nisse
; Tilgmann, Carola
LU
; Siuro, Jari
; Kervinen, Jukka
and Mikola, Leena
- publishing date
- 1992-02
- 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
- pmid:24186727
- 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
- 2024-01-04 01:04:49
@article{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).</p>}},
author = {{Sarkkinen, Paula and Kalkkinen, Nisse and Tilgmann, Carola and Siuro, Jari and Kervinen, Jukka and Mikola, Leena}},
issn = {{0032-0935}},
keywords = {{Aspartic proteinase; Cathepsin D; Endopeptidase; Hordeum (proteinase)}},
language = {{eng}},
number = {{3}},
pages = {{317--323}},
publisher = {{Springer}},
series = {{Planta}},
title = {{Aspartic proteinase from barley grains is related to mammalian lysosomal cathepsin D}},
url = {{http://dx.doi.org/10.1007/BF00195311}},
doi = {{10.1007/BF00195311}},
volume = {{186}},
year = {{1992}},
}