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Ribonuclease S‐peptide as a carrier in fusion proteins
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Kim, Jin-Soo | - |
dc.contributor.author | Raines, Ronald T. | - |
dc.date.accessioned | 2020-04-29T08:36:01Z | - |
dc.date.available | 2020-04-29T08:36:01Z | - |
dc.date.created | 2020-04-08 | - |
dc.date.created | 2020-04-08 | - |
dc.date.issued | 1993-03 | - |
dc.identifier.citation | Protein Science, Vol.2 No.3, pp.348-356 | - |
dc.identifier.issn | 0961-8368 | - |
dc.identifier.other | 95183 | - |
dc.identifier.uri | https://hdl.handle.net/10371/166251 | - |
dc.description.abstract | S-peptide (residues 1-20) and S-protein (residues 21-124) are the enzymatically inactive products of the limited digestion of ribonuclease A by subtilisin. S-peptide binds S-protein with high affinity to form ribonuclease S, which has full enzymatic activity. Recombinant DNA technology was used to produce a fusion protein having three parts: carrier, spacer, and target. The two carriers used were the first 15 residues of S-peptide (S15) and a mutant S15 in which Asp 14 had been changed to Asn (D14N S15). The spacer consisted of three proline residues and a four-residue sequence recognized by factor X(a) protease. The target was beta-galactosidase. The interaction between the S-peptide portion of the fusion protein and immobilized S-protein allowed for affinity purification of the fusion protein under denaturing (S15 as carrier) or nondenaturing (D14N S 1 5 as carrier) conditions. A sensitive method was developed to detect the fusion protein after sodium dodecyl sulfate-polyacrylamide gel electrophoresis by its ribonuclease activity following activation with S-protein. S-peptide has distinct advantages over existing carriers in fusion proteins in that it combines a small size (greater-than-or-equal-to 15 residues), a tunable affinity for ligand (K(d) greater-than-or-equal-to 10(-9) M), and a high sensitivity of detection (greater-than-or-equal-to 10(-16) mol in a gel). | - |
dc.language | 영어 | - |
dc.publisher | Cold Spring Harbor Laboratory Press | - |
dc.title | Ribonuclease S‐peptide as a carrier in fusion proteins | - |
dc.type | Article | - |
dc.contributor.AlternativeAuthor | 김진수 | - |
dc.identifier.doi | 10.1002/pro.5560020307 | - |
dc.citation.journaltitle | Protein Science | - |
dc.identifier.wosid | A1993KP46400007 | - |
dc.identifier.scopusid | 2-s2.0-0027512825 | - |
dc.citation.endpage | 356 | - |
dc.citation.number | 3 | - |
dc.citation.startpage | 348 | - |
dc.citation.volume | 2 | - |
dc.identifier.sci | A1993KP46400007 | - |
dc.description.isOpenAccess | N | - |
dc.contributor.affiliatedAuthor | Kim, Jin-Soo | - |
dc.type.docType | Article | - |
dc.description.journalClass | 1 | - |
dc.subject.keywordPlus | ESCHERICHIA-COLI | - |
dc.subject.keywordAuthor | ACTIVITY STAIN | - |
dc.subject.keywordAuthor | BIOTECHNOLOGY | - |
dc.subject.keywordAuthor | ENZYME ASSAY | - |
dc.subject.keywordAuthor | NONCOVALENT INTERACTION | - |
dc.subject.keywordAuthor | PROTEIN IMMOBILIZATION | - |
dc.subject.keywordAuthor | PROTEIN PURIFICATION | - |
dc.subject.keywordAuthor | SITE-DIRECTED MUTAGENESIS | - |
dc.subject.keywordAuthor | ZYMOGRAM | - |
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