TY - JOUR
T1 - Exploring the backbone dynamics of native spider silk proteins in Black Widow silk glands with solution-state NMR spectroscopy
AU - Xu, Dian
AU - Yarger, Jeffery
AU - Holland, Gregory P.
N1 - Funding Information:
This work was supported by grants from Department of Defense Air Force Office of Scientific Research (AFOSR) under Award No. FA9550-14-1-0014 , the Defense University Research Instrumentation Program (DURIP) under Award No. FA2386-12-1-3031 DURIP 12RSL231 and the National Science Foundation, Division of Materials Research (NSF-DMR) under Award No. DMR-1264801 . We thank Dr. Brian Cherry for help with NMR instrumentation and student training and Dr. Xu Wang for scientific discussion regarding protein solution-state NMR.
Copyright:
Copyright 2018 Elsevier B.V., All rights reserved.
PY - 2014/8/5
Y1 - 2014/8/5
N2 - Spider dragline silk is an outstanding biopolymer with a strength that exceeds steel by weight and a toughness greater than high-performance fibers like Kevlar. For this reason, understanding how a spider converts the gel-like, aqueous protein spinning dope within the major ampullate (MA) gland into a super fiber is of great importance for developing future biomaterials based on spider silk. In this work, the initial state of the silk proteins within Black Widow MA glands was probed with solution-state NMR spectroscopy. 15N relaxation parameters, T1, T2 and 15N-{ 1H} steady-state NOE were measured for twelve backbone environments at two spectrometer frequencies, 500 and 800 MHz. The NMR relaxation parameters extracted for all twelve environments are consistent with MA silk protein backbone dynamics on the fast sub-nanosecond timescale. Therefore, it is concluded that the repetitive core of spider MA proteins are in an unfolded, highly flexible state in the MA gland.
AB - Spider dragline silk is an outstanding biopolymer with a strength that exceeds steel by weight and a toughness greater than high-performance fibers like Kevlar. For this reason, understanding how a spider converts the gel-like, aqueous protein spinning dope within the major ampullate (MA) gland into a super fiber is of great importance for developing future biomaterials based on spider silk. In this work, the initial state of the silk proteins within Black Widow MA glands was probed with solution-state NMR spectroscopy. 15N relaxation parameters, T1, T2 and 15N-{ 1H} steady-state NOE were measured for twelve backbone environments at two spectrometer frequencies, 500 and 800 MHz. The NMR relaxation parameters extracted for all twelve environments are consistent with MA silk protein backbone dynamics on the fast sub-nanosecond timescale. Therefore, it is concluded that the repetitive core of spider MA proteins are in an unfolded, highly flexible state in the MA gland.
KW - Intrinsically disordered proteins
KW - NMR
KW - Spider silk protein
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U2 - 10.1016/j.polymer.2014.06.018
DO - 10.1016/j.polymer.2014.06.018
M3 - Article
AN - SCOPUS:84905509980
VL - 55
SP - 3879
EP - 3885
JO - Polymer (United Kingdom)
JF - Polymer (United Kingdom)
SN - 0032-3861
IS - 16
ER -