Comparative analysis of enzyme activities and mRNA levels of peptidyl prolyl cis/trans isomerases in various organs of wild type and Pin1-/- mice.
We investigated the enzyme activity of peptidyl prolyl cis/trans isomerases (PPIases) in brain, testis, lung, liver, and mouse embryonic fibroblasts (MEF) of Pin1+/+ and Pin1-/- mice. The aim of this study is to determine if other PPIases can substitute for the loss of Pin1 activity in Pin1-/- mice and what influence Pin1 depletion has on the activities of other PPIases members. The results show that high PPIase activities of Pin1 are found in organs that have the tendency to develop Pin1 knockout phenotypes and, therefore, provide for the first time an enzymological basis for these observations. Furthermore we determined the specific activity (k(cat)/K(M)) of endogenous Pin1 and found that it is strongly reduced as compared with the recombinant protein in all investigated organs. These results suggest that posttranslational modifications may influence the PPIase activity in vivo. The activities originating from cyclophilin and FKBP are not influenced by the Pin1 knockout, but a basal enzymatic activity towards phosphorylated substrates could be found in Pin1-/- lysates. Real time PCR experiments of all PPIases in different mouse organs and MEF of Pin1+/+ and Pin1-/- mice support the finding and reveal the specific expression profiles of PPIases in mice.
Fanghänel
Jörg
J
Center for Interdisciplinary Research, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi 981-8555, Japan.
Akiyama
Hirotada
H
Uchida
Chiyoko
C
Uchida
Takafumi
T
D000818:Animals; D001921:Brain / Q000201:enzymology; D021983:Cyclophilins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D005347:Fibroblasts / Q000201:enzymology; D020869:Gene Expression Profiling; D008099:Liver / Q000201:enzymology; D008168:Lung / Q000201:enzymology; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D019696:Peptidylprolyl Isomerase / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011487:Protein Conformation; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D022021:Tacrolimus Binding Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D013737:Testis / Q000201:enzymology; D014018:Tissue Distribution
D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D012333:RNA, Messenger; D021983:Cyclophilins; D022021:Tacrolimus Binding Proteins; D019696:Peptidylprolyl Isomerase; C000606850:Pin1 protein, mouse