Purification of a subtilisin inhibitor from black bean seeds

Seidl, Dinah S.; Abreu, Hugo; Jaffé, W. G.

doi:10.1016/0014-5793(78)80763-7

Cited by 19 publications

(8 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This finding also suggests a specificity for elastases (7), but only a weak "non-stoichiometric" inhibition of porcine pancreas elastase was found. Ala was also detected in the assumed P~ position of the Phaseolus inhibitor (14,15), which was a strong inhibitor of both subtilisin and leukocyte elastase, but the latter enzyme was not included in the present study.…”

Section: Discussionmentioning

confidence: 82%

“…Reactive sites in other inhibitors have been identified by cleavage at low pH with catalytic amounts of the enzyme inhibited (6,7,11,13,14,15). Attempts to identify a reactive site for subtilisin Carlsberg by this technique were, however, unsuccessful.…”

Section: Reactive Site(s)mentioning

confidence: 99%

“…pI 4.5 -6, which are not related to any of these two families of homologous inhibitors (2). Legume subtilisin inhibitors have, so far, been purified from seeds of black bean (Phaseolus vulgaris) (14) and cowpea (Vigna unguiculata) (19,20). Characterization of the Phaseolus inhibitor with respect to disulphide bridges and specificity (15) indicated relations to the "Streptomyces subtilisin inhibitor family" (7).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Subtilisin inhibitor from seeds of broad bean (vicia FABA); purification, amino acid sequence and specificity of inhibition

Svendsen¹,

Hejgaard²,

Chavan³

1984

Carlsberg Res. Commun.

View full text Add to dashboard Cite

A potent inhibitor of microbial serine proteases, including subtilisin, has been purified 1100-fold from seeds of broad bean (Vicia faba, cv. Kleine Thtiringer). Chymotrypsin and trypsin were not inhibited, but a weak "temporary" inhibition of pancreas elastase was observed. The preparation of pure inhibitor contained one major molecular form with a blocked N-terminal (MW -10,000, pl -4.8) and a minor form ( < 10%) with slightly lower MW and pl -5.0. About 80% of the amino acid sequence of the major form was determined by automatic Edman degradation of a cyanogen bromide fragment and tryptic peptides. The inhibitor is homologous with barley, potato and leech inhibitors of the "potato inhibitor I family" (36-56% of the amino acid residues in identical positions). Cleavage studies suggest one enzyme inhibitory site at an Ala-Asp bond.

show abstract

Section: Discussionmentioning

confidence: 82%

Section: Reactive Site(s)mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Subtilisin inhibitor from seeds of broad bean (vicia FABA); purification, amino acid sequence and specificity of inhibition

Svendsen¹,

Hejgaard²,

Chavan³

1984

Carlsberg Res. Commun.

View full text Add to dashboard Cite

show abstract

“…Other inhibitors of subtilisin were sub sequently purified from cereals and legumes. Subtilisin inhibitors are found in Hordeum vulgare (barley seeds) (17,191), Vigna unguiculata (cowpea) (180), Vicia faba (broad beans) (170), Phaseolus vulgaris (black beans) (160), Phaseolus angularis (adzuki bean) (192), Vigna radiata (mung bean) (89), Cicer arietinum (chick pea) (176) and Canavalia ensiformis (jack beans) (161). These inhibitors from legumes and cereals were first characterized by their high specificities toward microbial proteinases and their relative in effectiveness against serine proteinases from animals.…”

Section: Serine Proteinase Inhibitorsmentioning

confidence: 99%

Protease Inhibitors in Plants: Genes for Improving Defenses Against Insects and Pathogens

Ryan¹

1990

Annu. Rev. Phytopathol.

1,434

931

View full text Add to dashboard Cite

“…The inhibitors in black common bean seeds, a tryptophan content was determined by subtilisin inhlbitor was detected, purified and magnetic circular dichroism (2). The N-terminal partially characterized (1). residues of intact SI and enzyme cleaved SI It was shown that the subtilisin inhibitor was molecules were established by dansylation, selectively cleaved at two sites, one site by followed by acid hydrolysis and chromatosubtilisin and a second by bovine trypsin.…”

mentioning

confidence: 99%

Partial characterization of a subtilisin inhibitor from black bean seeds

Seidl

Abreu

Jaffé

1982

International Journal of Peptide and Protein Research

Self Cite

View full text Add to dashboard Cite

The subtilisin inhibitor (SI), isolated from black beans (Phaseolus vagaris) has a molecular weight of 8918 and it contains four half cystine residues. The N‐terminal residue is arginine and the C‐terminal is lysine. The subtilisin inhibiting site is composed of an Ala‐Leu or Ile bond. This site interacts in addition to subtilisin (S) with elastase from human leukocytes (HLE), both of which compete for the inhibitor. Also, HLE‐cleaved SI (SIHLE) is inactive against HLE as well as against S. It was shown that the inhibition of S by SI resembles a temporary inhibition, involving a very slow release of enzyme activity. The trypsin reactive site is a Lys‐Val bond, and its interaction with bovine trypsin is very weak. No complexes of SI or trypsin‐cleaved SI (SIT) with trypsin could be detected on cellulose‐acetate membrane electrophoresis, while SIT forms a clearly visible complex with S.

show abstract

Purification of a subtilisin inhibitor from black bean seeds

Cited by 19 publications

References 14 publications

Subtilisin inhibitor from seeds of broad bean (vicia FABA); purification, amino acid sequence and specificity of inhibition

Subtilisin inhibitor from seeds of broad bean (vicia FABA); purification, amino acid sequence and specificity of inhibition

Protease Inhibitors in Plants: Genes for Improving Defenses Against Insects and Pathogens

Partial characterization of a subtilisin inhibitor from black bean seeds

Contact Info

Product

Resources

About