Structure and dynamics of aromatic residues in spider silk: 2D carbon correlation NMR of dragline fibers.
The structure of aromatic residues in spider silk has remained relatively uncharacterized. This is primarily due to the relatively low abundance of aromatic residues in dragline spider silk. NMR characterization has been further hindered by the inability to (13)C-isotopically enrich these amino acids in the silk effectively. In the present contribution, it is shown that feeding spiders an aqueous solution of [U-(13)C/(15)N]-l-phenylalanine (13)C-enriches Tyr to a level of 20-30% in dragline spider silk. This enables the collection of 2D through-bond (13)C double quantum/single quantum (DQ/SQ) correlation spectra with the refocused INADEQUATE solid-state NMR pulse sequence. These 2D spectra provide the complete unambiguous assignment of the Tyr resonances in dragline silks from two spider species: N. clavipes and A. aurantia. Additionally, weak resonances are detected for Phe in A. aurantia dragline silk that is present at levels <1%. The conformation dependence of the C(alpha), C(beta), and carbonyl chemical shifts show that Tyr and Phe present in the GGX and GPGXX motifs in the spider silk proteins MaSp1 and MaSp2 are in disordered helical structures and not incorporated in the beta-sheet domains.
Izdebski
Thomas
T
Magnetic Resonance Research Center, Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona 85287-1604, USA.
Akhenblit
Paul
P
Jenkins
Janelle E
JE
Yarger
Jeffery L
JL
Holland
Gregory P
GP
D000818:Animals; D002247:Carbon Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010649:Phenylalanine / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D047011:Silk / Q000737:chemistry*; D013112:Spiders; D014443:Tyrosine / Q000737:chemistry*
D002247:Carbon Isotopes; D047011:Silk; D014443:Tyrosine; D010649:Phenylalanine