Dihydrofolate reductase mutant with exceptional resistance to methotrexate but not to trimetrexate

Pamela Pineda; Aaron Kanter; R. Scott McIvor; Stephen J. Benkovic; Andre Rosowsky; Carston R. Wagner

doi:10.1021/jm034057i

Dihydrofolate reductase mutant with exceptional resistance to methotrexate but not to trimetrexate

Pamela Pineda, Aaron Kanter, R. Scott McIvor, Stephen J. Benkovic, Andre Rosowsky, Carston R. Wagner

Research output: Contribution to journal › Article › peer-review

10 Scopus citations

Abstract

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 K_i 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.

Original language	English (US)
Pages (from-to)	2816-2818
Number of pages	3
Journal	Journal of medicinal chemistry
Volume	46
Issue number	14
DOIs	https://doi.org/10.1021/jm034057i
State	Published - Jul 3 2003

Access

10.1021/jm034057i

Fingerprint Dive into the research topics of 'Dihydrofolate reductase mutant with exceptional resistance to methotrexate but not to trimetrexate'. Together they form a unique fingerprint.

Cite this

@article{a244a2507e5b41faa403afccc50bb7f1,

title = "Dihydrofolate reductase mutant with exceptional resistance to methotrexate but not to trimetrexate",

abstract = "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.",

author = "Pamela Pineda and Aaron Kanter and McIvor, {R. Scott} and Benkovic, {Stephen J.} and Andre Rosowsky and Wagner, {Carston R.}",

year = "2003",

month = jul,

day = "3",

doi = "10.1021/jm034057i",

language = "English (US)",

volume = "46",

pages = "2816--2818",

journal = "Journal of Medicinal Chemistry",

issn = "0022-2623",

publisher = "American Chemical Society",

number = "14",

}

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

VL - 46

SP - 2816

EP - 2818

JO - Journal of Medicinal Chemistry

JF - Journal of Medicinal Chemistry

SN - 0022-2623

IS - 14

ER -

Dihydrofolate reductase mutant with exceptional resistance to methotrexate but not to trimetrexate

Abstract

Access

Other files and links

Cite this