Design and preparation of beta-sheet forming repetitive and block-copolymerized polypeptides.
The design and rapid construction of libraries of genes coding beta-sheet forming repetitive and block-copolymerized polypeptides bearing various C- and N-terminal sequences are described. The design was based on the assembly of DNA cassettes coding for the (GA)3GX amino acid sequence where the (GAGAGA) sequences would constitute the beta-strand units of a larger beta-sheet assembly. The edges of this beta-sheet would be functionalized by the turn-inducing amino acids (GX). The polypeptides were expressed in Escherichia coli using conventional vectors and were purified by Ni-nitriloacetic acid (NTA) chromatography. The correlation of polymer structure with molecular weight was investigated by gel electrophoresis and mass spectrometry. The monomer sequences and post-translational chemical modifications were found to influence the mobility of the polypeptides over the full range of polypeptide molecular weights while the electrophoretic mobility of lower molecular weight polypeptides was more susceptible to C- and N-termini polypeptide modifications.
Higashiya
Seiichiro
S
Department of Chemistry, University at Albany, SUNY, 1400 Washington Avenue, Albany, New York 12222, USA.
Topilina
Natalya I
NI
Ngo
Silvana C
SC
Zagorevskii
Dmitri
D
Welch
John T
JT
D000595:Amino Acid Sequence; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009571:Nitrilotriacetic Acid / Q000031:analogs & derivatives / Q000737:chemistry; D009942:Organometallic Compounds / Q000737:chemistry; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D017433:Protein Structure, Secondary; D020449:Repetitive Sequences, Amino Acid
D009942:Organometallic Compounds; D019151:Peptide Library; D010455:Peptides; C088321:nickel nitrilotriacetic acid; D009571:Nitrilotriacetic Acid