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Engineered disulfides improve mechanical properties of recombinant spider silk
Author(s) -
Grip S.,
Johansson J.,
Hedhammar M.
Publication year - 2009
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.111
Subject(s) - spider silk , silk , recombinant dna , biophysics , chemistry , disulfide bond , ultimate tensile strength , fiber , mutagenesis , polymer , materials science , biochemistry , mutation , biology , composite material , gene , organic chemistry
Abstract Nature's high‐performance polymer, spider silk, is composed of specific proteins, spidroins, which form solid fibers. So far, fibers made from recombinant spidroins have failed in replicating the extraordinary mechanical properties of the native material. A recombinant miniature spidroin consisting of four poly‐Ala/Gly‐rich tandem repeats and a nonrepetitive C‐terminal domain (4RepCT) can be isolated in physiological buffers and undergoes self assembly into macrofibers. Herein, we have made a first attempt to improve the mechanical properties of 4RepCT fibers by selective introduction of AA → CC mutations and by letting the fibers form under physiologically relevant redox conditions. Introduction of AA → CC mutations in the first poly‐Ala block in the miniature spidroin increases the stiffness and tensile strength without changes in ability to form fibers, or in fiber morphology. These improved mechanical properties correlate with degree of disulfide formation. AA → CC mutations in the forth poly‐Ala block, however, lead to premature aggregation of the protein, possibly due to disulfide bonding with a conserved Cys in the C‐terminal domain. Replacement of this Cys with a Ser, lowers thermal stability but does not interfere with dimerization, fiber morphology or tensile strength. These results show that mutagenesis of 4RepCT can reveal spidroin structure‐activity relationships and generate recombinant fibers with improved mechanical properties.