Document Detail

Substrate specificity of barley cysteine endoproteases EP-A and EP-B.
MedLine Citation:
PMID:  9576795     Owner:  NLM     Status:  MEDLINE    
The cysteine endoproteases (EP)-A and EP-B were purified from green barley (Hordeum vulgare L.) malt, and their identity was confirmed by N-terminal amino acid sequencing. EP-B cleavage sites in recombinant type-C hordein were determined by N-terminal amino acid sequencing of the cleavage products, and were used to design internally quenched, fluorogenic peptide substrates. Tetrapeptide substrates of the general formula 2-aminobenzoyl-P2-P1-P1'-P2'-tyrosine(NO2)-aspartic acid, in which cleavage occurs between P1 and P1', showed that the cysteine EPs preferred phenylalanine, leucine, or valine at P2. Arginine was preferred to glutamine at P1, whereas proline at P2, P1, or P1' greatly reduced substrate kinetic specificity. Enzyme cleavage of C hordein was mainly determined by the primary sequence at the cleavage site, because elongation of substrates, based on the C hordein sequence, did not make them more suitable substrates. Site-directed mutagenesis of C hordein, in which serine or proline replaced leucine, destroyed primary cleavage sites. EP-A and EP-B were both more active than papain, mostly because of their much lower Km values.
A Davy; I Svendsen; S O Sørensen; M Blom Sørensen; J Rouster; M Meldal; D J Simpson; V Cameron-Mills
Related Documents :
18358225 - High-throughput assays for sirtuin enzymes: a microfluidic mobility shift assay and a b...
1718435 - Substitution of phosphotyrosine for sulphotyrosine in biologically active peptides. enz...
2851585 - Porcine muscle prolyl endopeptidase and its endogenous substrates.
2966155 - Identification of the phosphorylation sites in early region 1a proteins of adenovirus t...
3941105 - High pressure liquid chromatography purification of up1 and up2, two related single-str...
1656175 - Recruitment of peripheral mononuclear cells by mammalian collagenase digests of type i ...
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Plant physiology     Volume:  117     ISSN:  0032-0889     ISO Abbreviation:  Plant Physiol.     Publication Date:  1998 May 
Date Detail:
Created Date:  1998-08-31     Completed Date:  1998-08-31     Revised Date:  2013-04-17    
Medline Journal Info:
Nlm Unique ID:  0401224     Medline TA:  Plant Physiol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  255-61     Citation Subset:  IM    
Carlsberg Research Laboratory, Department of Chemistry and Department of Physiology, Carlsberg Laboratory, Gammel Carlsbergvej 10, DK-2500 Valby, Denmark.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Amino Acid Sequence
Binding Sites
Chromogenic Compounds / metabolism
Cysteine Endopeptidases / genetics,  isolation & purification,  metabolism*
Enzyme Activation
Hordeum / enzymology*,  genetics
Molecular Sequence Data
Molecular Weight
Mutagenesis, Site-Directed
Plant Proteins / genetics,  metabolism
Substrate Specificity
Reg. No./Substance:
0/Chromogenic Compounds; 0/Plant Proteins; 8002-80-0/Glutens; EC 3.4.22.-/Cysteine Endopeptidases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Previous Document:  Cloning, expression in Escherichia coli, and characterization of Arabidopsis thaliana UMP/CMP kinase...
Next Document:  Isolation of the ornithine-delta-aminotransferase cDNA and effect of salt stress on its expression i...