TY - JOUR
T1 - Dihydrofolate reductase mutant with exceptional resistance to methotrexate but not to trimetrexate
AU - Pineda, Pamela
AU - Kanter, Aaron
AU - McIvor, R. Scott
AU - Benkovic, Stephen J.
AU - Rosowsky, Andre
AU - Wagner, Carston R.
PY - 2003/7/3
Y1 - 2003/7/3
N2 - Two double (F31A/F34A, I60A/L67G) and one quadruple (F31A/F34A/I60A/L67G) mutant murine dihydrofolate reductases were constructed and evaluated for their ability to impart antifolate resistance. Both I60A/L67G and F31A/F34A/I60A/L67G were found to be unstable and devoid of catalytic activity. The Ki values for F31A/F34A, methotrexate (MTX), bis-MTX, and PT-523 were found to be 10100-, 4410-, and 617-fold higher than the wild-type enzyme, respectively, but only 13.5-fold higher for trimetrexate (TMTX). These findings suggest that F31A/F34A could be used for gene therapy to render normal cells resistant to MTX but sensitive to TMTX.
AB - Two double (F31A/F34A, I60A/L67G) and one quadruple (F31A/F34A/I60A/L67G) mutant murine dihydrofolate reductases were constructed and evaluated for their ability to impart antifolate resistance. Both I60A/L67G and F31A/F34A/I60A/L67G were found to be unstable and devoid of catalytic activity. The Ki values for F31A/F34A, methotrexate (MTX), bis-MTX, and PT-523 were found to be 10100-, 4410-, and 617-fold higher than the wild-type enzyme, respectively, but only 13.5-fold higher for trimetrexate (TMTX). These findings suggest that F31A/F34A could be used for gene therapy to render normal cells resistant to MTX but sensitive to TMTX.
UR - http://www.scopus.com/inward/record.url?scp=0038460886&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0038460886&partnerID=8YFLogxK
U2 - 10.1021/jm034057i
DO - 10.1021/jm034057i
M3 - Article
C2 - 12825924
AN - SCOPUS:0038460886
SN - 0022-2623
VL - 46
SP - 2816
EP - 2818
JO - Journal of medicinal chemistry
JF - Journal of medicinal chemistry
IS - 14
ER -