Accelerated chemical synthesis of peptides and small proteins

Miranda, LP and Alewood, PF (1999) Accelerated chemical synthesis of peptides and small proteins. Proc. Natl Acad. Sci, 96 4: 1181-1186. doi:10.1073/pnas.96.4.1181

Author Miranda, LP
Alewood, PF
Title Accelerated chemical synthesis of peptides and small proteins
Journal name Proc. Natl Acad. Sci   Check publisher's open access policy
ISSN 0027-8424
Publication date 1999
Sub-type Article (original research)
DOI 10.1073/pnas.96.4.1181
Open Access Status Not Open Access
Volume 96
Issue 4
Start page 1181
End page 1186
Total pages 6
Place of publication USA
Publisher National Academy of Science
Collection year 1999
Language eng
Subject C1
250301 Organic Chemical Synthesis
780103 Chemical sciences
Abstract The chemical synthesis of peptides and small proteins is a powerful complementary strategy to recombinant protein overexpression and is widely used in structural biology, immunology, protein engineering, and biomedical research. Despite considerable improvements in the fidelity of peptide chain assembly, side-chain protection, and postsynthesis analysis, a limiting factor in accessing polypeptides containing greater than 50 residues remains the time taken for chain assembly. The ultimate goal of this work is to establish highly efficient chemical procedures that achieve chain-assembly rates of approximately 10-15 residues per hour, thus underpinning the rapid chemical synthesis of long polypeptides and proteins, including cytokines, growth factors, protein domains, and small enzymes, Here we report Boc chemistry that employs O-(7-azabenzo-triazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU)/dimethyl sulfoxide in situ neutralization as the coupling agent and incorporates a protected amino acid residue every 5 min to produce peptides of good quality. This rapid coupling chemistry was successfully demonstrated by synthesizing several small to medium peptides, including the difficult C-terminal sequence of HIV-1 proteinase (residues 81-99); fragment 65-74 of the acyl carrier protein; conotoxin PnIA(A10L), a potent neuronal nicotinic receptor antagonist; and the proinflammatory chemotactic protein CP10, an 88-residue protein, by means of native chemical ligation, The benefits of this approach include enhanced ability to identify and characterize difficult couplings, rapid access to peptides for biological and structure-activity studies, and accelerated synthesis of tailored large peptide segments (<50 residues) for use in chemoselective ligation methods.
Keyword Chemistry, Organic
Solid-phase Synthesis
Amino-acid Fluorides
Hiv-1 Protease
Difficult Sequences
Coupling Reagents
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collection: Institute for Molecular Bioscience - Publications
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Citation counts: TR Web of Science Citation Count  Cited 62 times in Thomson Reuters Web of Science Article | Citations
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Created: Tue, 10 Jun 2008, 15:17:18 EST