Specific glutamine and asparagine residues of gamma-S crystallin are resistant to in vivo deamidation.
It has been hypothesized that resistance to nonenzymatic deamidation of asparagine and glutamine residues may be an important determinant of protein stability in vivo. As a test of this hypothesis, we analyzed the central region of old human lenses, which contain proteins such as gamma-S crystallin that were synthesized during the fetal-embryonic periods of development. Total protein from the fetal-embryonic region of old human lenses was digested with trypsin, followed by resolution of tryptic fragments containing amidated and deamidated forms using high pressure liquid chromatography-reverse phase chromatography together with synthetic peptide standards and mass spectral analysis. The results demonstrate no detectable deamidation of glutamine 92, glutamine 96, asparagine 143, and glutamine 170 from gamma-S crystallin from old human lenses, consistent with the hypothesis that very long-lived proteins can contain asparagine and glutamine residues that are extremely resistant to in vivo deamidation.
Takemoto
L
L
Division of Biology, Ackert Hall, Kansas State University, Manhattan, Kansas 66506, USA. takemlj@ksu.edu
Boyle
D
D
D000368:Aged; D000595:Amino Acid Sequence; D001216:Asparagine / Q000378:metabolism*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D003459:Crystallins / Q000378:metabolism*; D005973:Glutamine / Q000378:metabolism*; D006801:Humans; D008875:Middle Aged; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010449:Peptide Mapping; D013329:Structure-Activity Relationship
D003459:Crystallins; D005973:Glutamine; D001216:Asparagine