Flexibility in the solution structure of human tropoelastin.
We investigated the flexibility of full-length tropoelastin in solution by using far- and near-ultraviolet circular dichroism (UV CD) and fluorescence spectroscopy to probe for structural flexibility and residue mobility within secondary and tertiary features of the monomer. Fluorescence spectroscopy revealed the presence of exposed hydrophobicity through the binding of the hydrophobic probe 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonate (bis-ANS), which demonstrates that hydrophobic regions form clusters and are not confined to a molecular core. Near-UV CD indicated substantial mobility of aromatic residues. Structural prediction programs (PONDR, DisEMBL, and Globplot version 2.0) estimated 75 +/- 2% disorder in the tertiary structure of tropoelastin on the basis of primary sequence information. A single-site substitution of Trp for Gln (Q513W) at the tropoelastin domain 25-26 interface facilitated fluorescence spectroscopy for revealing that this region is exposed to solvent. Polarization anisotropy demonstrated substantial flexibility of W513 and little change upon denaturation of the monomer with guanidine hydrochloride. Comparable movement was found for native sequence aromatic residues in the presence of glycosaminoglycans and trifluoroethanol. These data prove the intrinsic flexibility of specific residues and adjacent sequences in any native conformation(s) they may take. This study is the first characterization of the level of mobility in defined regions of the full-length tropoelastin monomer and provides direct evidence for regions of flexible structure in tropoelastin.
Muiznieks
Lisa D
LD
School of Molecular and Microbial Biosciences, The University of Sydney, Sydney, Australia 2006.
Weiss
Anthony S
AS
D000595:Amino Acid Sequence; D001483:Base Sequence; D002942:Circular Dichroism; D017931:DNA Primers; D006801:Humans; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D014333:Tropoelastin / Q000737:chemistry*
D017931:DNA Primers; D014333:Tropoelastin