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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
12
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pubmed:dateCreated |
2001-6-6
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pubmed:abstractText |
We have completed the total chemical synthesis of cytochrome b562 and an axial ligand analogue, [SeMet(7)]cyt b562, by thioester-mediated chemical ligation of unprotected peptide segments. A novel auxiliary-mediated native chemical ligation that enables peptide ligation to be applied to protein sequences lacking cysteine was used. A cleavable thiol-containing auxiliary group, 1-phenyl-2-mercaptoethyl, was added to the alpha-amino group of one peptide segment to facilitate amide bond-forming ligation. The amine-linked 1-phenyl-2-mercaptoethyl auxiliary was stable to anhydrous hydrogen fluoride used to cleave and deprotect peptides after solid-phase peptide synthesis. Following native chemical ligation with a thioester-containing segment, the auxiliary group was cleanly removed from the newly formed amide bond by treatment with anhydrous hydrogen fluoride, yielding a full-length unmodified polypeptide product. The resulting polypeptide was reconstituted with heme and folded to form the functional protein molecule. Synthetic wild-type cyt b562 exhibited spectroscopic and electrochemical properties identical to the recombinant protein, whereas the engineered [SeMet(7)]cyt b562 analogue protein was spectroscopically and functionally distinct, with a reduction potential shifted by approximately 45 mV. The use of the 1-phenyl-2-mercaptoethyl removable auxiliary reported here will greatly expand the applicability of total protein synthesis by native chemical ligation of unprotected peptide segments.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-10353818,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-10359755,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-10393541,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-10468563,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-10600727,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-10685043,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-10821680,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-10891251,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-10966479,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-11087311,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-11101297,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-11170453,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-11456561,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-1310985,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-1478777,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-1566069,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-2560194,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-4399869,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-5330963,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-5336964,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-7723042,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-7973629,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-8332196,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-8900071,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-9353726,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11390992-9720266
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Jun
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pubmed:issn |
0027-8424
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
5
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pubmed:volume |
98
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
6554-9
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
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pubmed:year |
2001
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pubmed:articleTitle |
Total synthesis of cytochrome b562 by native chemical ligation using a removable auxiliary.
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pubmed:affiliation |
Gryphon Sciences, South San Francisco, CA 94080; and Department of Chemistry, Occidental College, Los Angeles, CA 90041, USA. dlow@gryphonsci.com
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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