Protein L-isoaspartyl methyltransferase catalyzes in vivo racemization of Aspartate-25 in mammalian histone H2B.
Protein L-isoaspartyl methyltransferase (PIMT) has been implicated in the repair or metabolism of proteins containing atypical L-isoaspartyl peptide bonds. The repair hypothesis is supported by previous studies demonstrating in vitro repair of isoaspartyl peptides via formation of a succinimide intermediate. Utilization of this mechanism in vivo predicts that PIMT modification sites should exhibit significant racemization as a side reaction to the main repair pathway. We therefore studied the D/L ratio of aspartic acid at specific sites in histone H2B, a known target of PIMT in vivo. Using H2B from canine brain, we found that Asp25 (the major PIMT target site in H2B) was significantly racemized, exhibiting d/l ratios as high as 0.12, whereas Asp51, a comparison site, exhibited negligible racemization (D/L < or = 0.01). Racemization of Asp25 was independent of animal age over the range of 2-15 years. Using H2B from 2-3-week mouse brain, we found a similar D/L ratio (0.14) at Asp25 in wild type mice, but substantially less racemization (D/L = 0.035) at Asp25 in PIMT-deficient mice. These findings suggest that PIMT functions in the repair, rather than the metabolic turnover, of isoaspartyl proteins in vivo. Because PIMT has numerous substrates in cells, these findings also suggest that D-aspartate may be more common in cellular proteins than hitherto imagined and that its occurrence, in some proteins at least, is independent of animal age.
Young
Glen W
GW
Department of Molecular Biology and Biochemistry, University of California, Irvine, California 92697, USA.
Hoofring
Sarah A
SA
Mamula
Mark J
MJ
Doyle
Hester A
HA
Bunick
Gerard J
GJ
Hu
Yonglin
Y
Aswad
Dana W
DW
D000595:Amino Acid Sequence; D000818:Animals; D001224:Aspartic Acid / Q000737:chemistry*; D001921:Brain / Q000378:metabolism; D002384:Catalysis; D002843:Chromatin / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D004285:Dogs; D006657:Histones / Q000737:chemistry* / Q000378:metabolism*; D008956:Models, Chemical; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D026601:Protein D-Aspartate-L-Isoaspartate Methyltransferase / Q000378:metabolism / Q000502:physiology*; D017386:Sequence Homology, Amino Acid; D013237:Stereoisomerism; D013997:Time Factors
D002843:Chromatin; D006657:Histones; D010455:Peptides; D001224:Aspartic Acid; D026601:Protein D-Aspartate-L-Isoaspartate Methyltransferase