Original language | English |
---|---|
Pages (from-to) | 365-368 |
Number of pages | 4 |
Journal | Advances in Experimental Medicine and Biology |
Volume | 431 |
DOIs | |
State | Published - 1998 |
Funding
Funders | Funder number |
---|---|
Medical Research Council Canada |
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In: Advances in Experimental Medicine and Biology, Vol. 431, 1998, p. 365-368.
Research output: Contribution to journal › Article › peer-review
TY - JOUR
T1 - Adenosine-induced preconditioning of rat neuronal cultures against ischemia-reperfusion injury
AU - Reshef, A.
AU - Sperling, O.
AU - Zoref-Shani, E.
N1 - Funding Information: C.F. Cuncic and F.F. Snyder, Departments of Medical Biochemistry and Medical Genetics, University of Calgary, Calgary, Alberta, Canada Objectives We previously characterized IMP dehydrogenase from a mycophenolic acid resistant cell line as having two substitutions, Thr333Ile and Ser351Tyr. An independent study of the consequence of these two residues was undertaken. Design The single mutants, Ile333 and Tyr351 were produced by site directed mutagenesis. Control, double and single mutants were subcloned and expressed in pMal and purified by affinity chromatography for kinetic analysis. Results Comparative Vmax studies revealed the Ile333 substitution was responsible for a 10-fold reduction in activity. Tyr351 produced the greatest increase in Km for IMP, 6-fold, compared to 1.9-fold for Ile333, and 2.8-fold for Ile333fryr351. Ile333 alone produced a 10.6-fold increase in Km for NAD, Tyr351, 5.4-fold and Ile333fryr351 4.4-fold. Ile333 also produced the greatest increase in Ki for XMP, 6.7-fold. Inhibition constants for mycophenolic acid with recombinant IMP dehydrogenase were consistent with previous findings with Ki's of 17.6 and 2950 nM for control and Ile333fryr351 respectively. This change was almost entirely due to the Ile333 mutation and not Tyr351, having Ki's of 2300 and 23 nM respectively. Conclusion Thr333 has an important role in IMP dehydrogenase sensitivity to mycophenolic acid as alteration to Ile results in a 140-increase in Ki. Supported by the Medical Research Council of Canada.
PY - 1998
Y1 - 1998
UR - http://www.scopus.com/inward/record.url?scp=0031863423&partnerID=8YFLogxK
U2 - 10.1007/978-1-4615-5381-6_72
DO - 10.1007/978-1-4615-5381-6_72
M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???
AN - SCOPUS:0031863423
SN - 0065-2598
VL - 431
SP - 365
EP - 368
JO - Advances in Experimental Medicine and Biology
JF - Advances in Experimental Medicine and Biology
ER -