Purification and characterization of a chymosinlike protease from the gastric mucosa of harp seal ( Pagophilus groenlandicus )
Four zymogens of acidic proteases A, B, C, and D were isolated from the gastric mucosa of harp seals by ion-exchange chromatography on a diethylaminoethyl-Sephadex A-50 column. The major zymogens were A and C, and the ratio of zymogen A to zymogen C was greater in extracts from 1-week-old animals th...
Published in: | Canadian Journal of Biochemistry and Cell Biology |
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1984
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crcansciencepubl:10.1139/o84-091 2023-12-17T10:31:18+01:00 Purification and characterization of a chymosinlike protease from the gastric mucosa of harp seal ( Pagophilus groenlandicus ) Shamsuzzaman, K. Haard, N. F. 1984 http://dx.doi.org/10.1139/o84-091 http://www.nrcresearchpress.com/doi/pdf/10.1139/o84-091 en eng Canadian Science Publishing http://www.nrcresearchpress.com/page/about/CorporateTextAndDataMining Canadian Journal of Biochemistry and Cell Biology volume 62, issue 8, page 699-708 ISSN 0714-7511 General Medicine journal-article 1984 crcansciencepubl https://doi.org/10.1139/o84-091 2023-11-19T13:38:21Z Four zymogens of acidic proteases A, B, C, and D were isolated from the gastric mucosa of harp seals by ion-exchange chromatography on a diethylaminoethyl-Sephadex A-50 column. The major zymogens were A and C, and the ratio of zymogen A to zymogen C was greater in extracts from 1-week-old animals than in extracts from adult animals. Zymogens A and C were further purified by affinity chromatography using carbobenzoxy-D-phenylalaninetriethylene tetramine Sepharose and gel filtration on a Sephadex G-100 column. Certain physical and catalytic properties of proteases A and C were compared with those of calf chymosin (EC 3.4.23.4) and porcine pepsin (EC 3.4.23.1). Zymogen C and the corresponding enzyme were homogeneous on analytical polyacrylamide gel electrophoresis. Zymogen A was homogeneous as judged by sodium dodecyl sulphate (SDS) – polyacrylamide gel electrophoresis and high performance liquid chromatography, but was heterogenous by polyacrylamide gel electrophoresis at pH 8.3. Zymogens A and C had molecular weights of 33 800 and 44 000, respectively, as estimated by SDS–polyacrylamide gel electrophoresis. Protease A had an isoelectric point of 4.90. Protease A was similar to calf chymosin with respect to several criteria. It had a higher ratio of milk-clotting to proteolytic activity than those of seal protease C and porcine pepsin and had a pH optimum of 2.2–3.5 for hemoglobin hydrolysis. It did not inactivate ribonuclease, had very low activity on N-acetyl-L-phenylalanyl-3,5-diiodo-L-tyrosine and lost activity in 6 M urea. These results indicate protease A is chymosinlike. Article in Journal/Newspaper Harp Seal Pagophilus groenlandicus Canadian Science Publishing (via Crossref) Canadian Journal of Biochemistry and Cell Biology 62 8 699 708 |
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Canadian Science Publishing (via Crossref) |
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English |
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General Medicine |
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General Medicine Shamsuzzaman, K. Haard, N. F. Purification and characterization of a chymosinlike protease from the gastric mucosa of harp seal ( Pagophilus groenlandicus ) |
topic_facet |
General Medicine |
description |
Four zymogens of acidic proteases A, B, C, and D were isolated from the gastric mucosa of harp seals by ion-exchange chromatography on a diethylaminoethyl-Sephadex A-50 column. The major zymogens were A and C, and the ratio of zymogen A to zymogen C was greater in extracts from 1-week-old animals than in extracts from adult animals. Zymogens A and C were further purified by affinity chromatography using carbobenzoxy-D-phenylalaninetriethylene tetramine Sepharose and gel filtration on a Sephadex G-100 column. Certain physical and catalytic properties of proteases A and C were compared with those of calf chymosin (EC 3.4.23.4) and porcine pepsin (EC 3.4.23.1). Zymogen C and the corresponding enzyme were homogeneous on analytical polyacrylamide gel electrophoresis. Zymogen A was homogeneous as judged by sodium dodecyl sulphate (SDS) – polyacrylamide gel electrophoresis and high performance liquid chromatography, but was heterogenous by polyacrylamide gel electrophoresis at pH 8.3. Zymogens A and C had molecular weights of 33 800 and 44 000, respectively, as estimated by SDS–polyacrylamide gel electrophoresis. Protease A had an isoelectric point of 4.90. Protease A was similar to calf chymosin with respect to several criteria. It had a higher ratio of milk-clotting to proteolytic activity than those of seal protease C and porcine pepsin and had a pH optimum of 2.2–3.5 for hemoglobin hydrolysis. It did not inactivate ribonuclease, had very low activity on N-acetyl-L-phenylalanyl-3,5-diiodo-L-tyrosine and lost activity in 6 M urea. These results indicate protease A is chymosinlike. |
format |
Article in Journal/Newspaper |
author |
Shamsuzzaman, K. Haard, N. F. |
author_facet |
Shamsuzzaman, K. Haard, N. F. |
author_sort |
Shamsuzzaman, K. |
title |
Purification and characterization of a chymosinlike protease from the gastric mucosa of harp seal ( Pagophilus groenlandicus ) |
title_short |
Purification and characterization of a chymosinlike protease from the gastric mucosa of harp seal ( Pagophilus groenlandicus ) |
title_full |
Purification and characterization of a chymosinlike protease from the gastric mucosa of harp seal ( Pagophilus groenlandicus ) |
title_fullStr |
Purification and characterization of a chymosinlike protease from the gastric mucosa of harp seal ( Pagophilus groenlandicus ) |
title_full_unstemmed |
Purification and characterization of a chymosinlike protease from the gastric mucosa of harp seal ( Pagophilus groenlandicus ) |
title_sort |
purification and characterization of a chymosinlike protease from the gastric mucosa of harp seal ( pagophilus groenlandicus ) |
publisher |
Canadian Science Publishing |
publishDate |
1984 |
url |
http://dx.doi.org/10.1139/o84-091 http://www.nrcresearchpress.com/doi/pdf/10.1139/o84-091 |
genre |
Harp Seal Pagophilus groenlandicus |
genre_facet |
Harp Seal Pagophilus groenlandicus |
op_source |
Canadian Journal of Biochemistry and Cell Biology volume 62, issue 8, page 699-708 ISSN 0714-7511 |
op_rights |
http://www.nrcresearchpress.com/page/about/CorporateTextAndDataMining |
op_doi |
https://doi.org/10.1139/o84-091 |
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Canadian Journal of Biochemistry and Cell Biology |
container_volume |
62 |
container_issue |
8 |
container_start_page |
699 |
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708 |
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1785584526903214080 |