Elevated Amounts of Methotrexate-binding Protein, Different from Normal Dihydrofolate Reductase, in a Petunia MTX(R)-cell Line

Overview

Journal Plant Mol Biol

Date 2013 Dec 5

PMID 24302527

Citations 1

Authors

R Barg

M Perl

J S Beckmann

Affiliations

Soon will be listed here.

Abstract

A petunia cell line, 1ECB, was previously isolated by the stepwise selection procedure, for resistance to methotrexate (MTX), an antimetabolite for the enzyme dihydrofolate reductase (DHFR). Using ammonium sulfate precipitates of cell lysates of cell line 1ECB and its parental cell line (WT), it was found that the mutant has an increase of 400 fold in (3)H-MTX binding capacity and a decrease in the affinity for MTX binding, at two orders of magnitude, in comparison with the WT. In addition, the DHFR specific activity in the mutant increased only moderately (5- to 10-fold), this activity is extremely sensitive to MTX inhibition as compared to the WT. It is evident that the MTX resistance of line 1ECB results mainly from overproduction of an MTX-binding protein which differs from the WT DHFR by four biochemical criteria. This protein may serve as a trap for the excess amounts of MTX to which the cells are exposed.

Citing Articles

Reduced intracellular content of methotrexate in an isolated MTX-resistant cell line of Nicotiana plumbaginifolia.

Barg R, Shabtai S Plant Cell Rep. 2013; 10(5):269-72.

PMID: 24221594 DOI: 10.1007/BF00232573.

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