A description of the nature of the transition state structure for phosphoryl transfer in the cAPK reaction requires a measurement of the primary 180 isotope effect at the serine hydroxyl acceptor. Since it is difficult to obtain primary 180 isotope effect directly, the 15N/1 4N ratio of the a-amine of the C-terminal glycine in the peptide Leu Arg-Lys-Ala-Ser-Leu-Gly (when serine is phosphorylated) was used to represent on the phosphorylation at serine. 15N Glycine, ' 4N-Glycine and 180 serine were synthesized and used to synthesize two peptides, one containing 1 80-serine/' 5 N glycine and second 1 60-serine/1 4N-glycine. Methods were developed for hydrolyzing the peptides and quantitatively isolating glycine. Partitioning results suggest that catalytic rate was slow compare to substrate dissociation. The 180 primary isotope effect will be determined in the near future using the method developed herein.
| Identifer | oai:union.ndltd.org:unt.edu/info:ark/67531/metadc500775 |
| Date | 12 1900 |
| Creators | Chen, Gang, 1963- |
| Contributors | Norton, S. J., Cook, Paul,1944-, Wu, Edward Ming-chi, 1938- |
| Publisher | University of North Texas |
| Source Sets | University of North Texas |
| Language | English |
| Detected Language | English |
| Type | Thesis or Dissertation |
| Format | v, 43 leaves : ill., Text |
| Rights | Public, Chen, Gang, 1963-, Copyright, Copyright is held by the author, unless otherwise noted. All rights reserved. |
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