Amino acid analysis of spider dragline silk using 1H NMR

Xiangyan Shi, Gregory P. Holland, Jeffery Yarger

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

The amino acid composition of Nephila clavipes dragline silk fiber was determined by conducting 1H nuclear magnetic resonance (NMR) spectroscopy experiments on acid-hydrolyzed material. N. clavipes dragline silk was found to consist of 43.0 ± 0.6% Gly, 29.3 ± 0.2% Ala, 9.1 ± 0.1% Glx, 4.0 ± 0.1% Leu, 3.3 ± 0.1% Tyr, 3.4 ± 0.2% Ser, 2.7 ± 0.1% Pro, 2.1 ± 0.1% Arg, 1.07 ± 0.05% Asx, 0.96 ± 0.05% Val, 0.48 ± 0.03% Thr, 0.35 ± 0.03% Phe, and 0.28 ± 0.03% Ile. Compared with standard chromatography-based amino acid analysis (AAA), the chemical resolution of NMR allows for an amino acid solution to be characterized without separation and is shown to provide considerably higher precision. This allows for more accurate statistics on the variability of amino acids in spider dragline silk. In general, this 1H NMR AAA technique is applicable to a large range of proteins and peptides for precise composition characterization, especially when the precise content of a minor component is critical and relatively large amounts of sample are available (microgram to milligram quantities).

Original languageEnglish (US)
Pages (from-to)150-157
Number of pages8
JournalAnalytical Biochemistry
Volume440
Issue number2
DOIs
Publication statusPublished - 2013

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Keywords

  • H NMR
  • Amino acid analysis
  • Nephila clavipes dragline silk
  • Spider silk

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Cell Biology

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