One- and two-dimensional NMR spectral analysis of the consequences of single amino acid replacements in proteins.
The traditional approach of using homologous sequences to elucidate the role of specific amino acid residues in protein structure and function becomes more meaningful as the number of differences is minimized, with the limit being alteration of a single residue. For small proteins in solution, NMR spectroscopy offers a means of obtaining detailed information about each residue and its response to a given change in the protein sequence. Extraction of this information has been aided by recent progress in spectrometer technology (higher magnetic fields, more sensitive signal detection, more sophisticated computers) and experimental strategies (new NMR pulse sequences including multiple-quantum and two-dimensional NMR methods). The set of avian ovomucoid third domains, which consists of the third domain proper plus a short leader (connecting peptide) and has a maximum of 56 amino acid residues, offers an attractive system for developing experimental methods for investigating sequence-structure and structure-function relationships in proteins. Our NMR results provide examples of sequence effects on pKa' values, average conformation, and internal motion of amino acid side chains.
J L
JL
Markley
D H
DH
Croll
R
R
Krishnamoorthi
G
G
Ortiz-Polo
W M
WM
Westler
W C
WC
Bogard
M
M
Laskowski
D000595:Amino Acid Sequence; D000818:Animals; D001717:Birds; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D010059:Ovomucin; D011487:Protein Conformation; D014443:Tyrosine
D010059:Ovomucin; D014443:Tyrosine