TY - JOUR
T1 - Amino acid analysis of spider dragline silk using 1H NMR
AU - Shi, Xiangyan
AU - Holland, Gregory P.
AU - Yarger, Jeffery
N1 - Funding Information:
We thank Professor Randy Lewis and Dr. Justin Jones (currently working at Utah State University) for help with collecting AAA data on the hydrolyzed silk samples. We also thank Dr. Brian Cherry for help with NMR instrumentation, student training, and scientific discussion. This work is supported by grants from the Department of Defense Air Force Office of Scientific Research under Award FA9550-10-1-0275, Defense University Research Instrumentation Program (DURIP) under Award No. FA2386-12-1-3031 DURIP 12RSL231, and the National Science Foundation, Division of Materials Research, under Award DMR-1264801.
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2013
Y1 - 2013
N2 - 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).
AB - 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).
KW - Amino acid analysis
KW - Nephila clavipes dragline silk
KW - Spider silk
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U2 - 10.1016/j.ab.2013.05.006
DO - 10.1016/j.ab.2013.05.006
M3 - Article
C2 - 23727559
AN - SCOPUS:84884939154
SN - 0003-2697
VL - 440
SP - 150
EP - 157
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -