title: Folding recombinant spider-silk in H2 O: Effect of osmolytes on the solution conformation of a 15-repeat spider-silk mimetic.
abstract: The folding of a recombinant spider silk protein-polymer in the presence of the tri-methylamine osmolytes TMANO and Betaine in 80% H2 O is reported. Circular dichroism measurements (CD) reveal an increase in α-helical secondary structure with increasing osmolyte concentrations, as determined by an increase in ellipticity at 222 nm. Consistent with this observation, the signal for random coil sampling, observed at 205 nm, is greatly reduced with increasing trimethylamine. Fluorescence spectra of a single tyrosine positioned within the conserved 33-amino acid repeat primary sequence (of the spider-silk mimetic) complements the conformational changes observed by CD. Importantly, there is a correlation between the number of Alkyl-groups (CH3 -) on the amine of the osmolyte and enhanced helicity of the 15-repeat silk-mimetic for the osmolytes tested, ie TMANO, Betaine, Sarcosine and Glycine. These preliminary results are applicable to storing and processing recombinant silk sequences in H2 O, an important mile-stone for widespread use of recombinant silk polymers.
authors.affiliation: Department of Chemistry and Biochemistry Queens College, CUNY, Flushing, NY, 11367. drdale.mclach@gmail.com.
authors.forename: Glendon D
authors.initials: GD
authors.lastname: McLachlan
authors.affiliation: Department of Chemistry and Biochemistry Queens College, CUNY, Flushing, NY, 11367.
authors.forename: Babak
authors.initials: B
authors.lastname: Gandjian
authors.affiliation: Department of Chemistry and Biochemistry Queens College, CUNY, Flushing, NY, 11367.
authors.forename: Hind
authors.initials: H
authors.lastname: Alhumaidan
mesh_terms: D000818:Animals; D057786:Peptidomimetics / Q000737:chemistry*; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D047011:Silk / Q000737:chemistry* / Q000235:genetics; D013112:Spiders; D014867:Water
chemical_list: D057786:Peptidomimetics; D011994:Recombinant Proteins; D047011:Silk; D014867:Water
keywords: TMANO; betaine; circular dichroism; osmolytes; protein folding; sarcosine; spider-silk mimetic; taurine