Amino acid analysis of spider dragline silk using H-1 NMR

作者：Shi Xiangyan; Holland Gregory P^*; Yarger Jeffery L

来源：Analytical Biochemistry, 2013, 440(2): 150-157.

DOI：10.1016/j.ab.2013.05.006

• Nephila clavipes dragline silk
• H-1 NMR
• Amino acid analysis
• Spider silk

The amino acid composition of Nephila clavipes dragline silk fiber was determined by conducting H-1 nuclear magnetic resonance (NMR) spectroscopy experiments on acid-hydrolyzed material. N. davipes 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% Mg, 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 H-1 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). Published by Elsevier Inc.

• 出版日期2013-9-15