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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
5
|
| pubmed:dateCreated |
1981-6-13
|
| pubmed:abstractText |
While trypsin can catalyze resynthesis of the peptide bond between fragments in the noncovalent complex of nuclease-T-(6-48) and nuclease-T-(49-149), this reaction leads to excision of Lys 49 and formation of inactive [des Lys 49]-nuclease-(6-149). To provide a method for making active active covalent semisynthetic nuclease, we chemically synthesized the fragment of residues 6 to 49 in which lysine 48 was replaced by glycine. This peptide was made using the recently described solid phase support, 4-(oxymethyl)phenylacetamidomethyl-polystyrene. The resultant crude polypeptide exhibited 30-50% of native nuclease-T enzymatic activity when added to native nuclease-T-(50-149). When the non-covalent complex formed by native nuclease-T-(50-149) and a 10-fold molar excess of [Gly 48]synthetic-(6-49) was equilibrated with trypsin in 90% glycerol, an increase in enzymatic activity from 8 to 32% (versus nuclease) was observed. Simultaneously, approximately 20% conversion of nuclease-T-(50-149) to nuclease-molecular weight material was observed by gel electrophoretic analysis. These data indicate that a covalent semisynthetic species is formed with activity about equal to that of native nuclease. The results confirm the importance of loop integrity on catalytic site organization. The Gly-48-containing fragment system defined above can allow preparation of semisynthetic nuclease sequence analogs.
|
| pubmed:grant | |
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Nov
|
| pubmed:issn |
0367-8377
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
16
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
433-9
|
| pubmed:dateRevised |
2007-11-14
|
| pubmed:meshHeading | |
| pubmed:year |
1980
|
| pubmed:articleTitle |
Enzyme-catalyzed formation of semisynthetic staphylococcal nuclease using a new synthetic fragment, [48-glycine]synthetic-(6-49).
|
| pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
|