Heterogeneity of methotrexate binding in human colon tumor cells.
[3H]-methotrexate binding at pH 5.0 and pH 7.2 by the cytosol of tumor tissues and the surrounding normal areas of the gastrointestinal tract of patients suffering from colon or gastric cancer has been used to identify in these cells the presence of a binder of methotrexate having low-affinity for this drug in addition to the enzyme dihydrofolate reductase. Scatchard analysis of the [3H]-methotrexate binding data by a colon tumor sample also reveals that there are two binders of this drug present in the cytosol of these cells. The association constant (Kass) for one binder of methotrexate is = 5.6 x 10(7) M-1 while the Kass for the second binder is = 1.0 x 10(6) M-1. The two binders do not differ very much in their apparent molecular weight. Upon isoelectric focusing, the tumor cell cytosol resolves into 4 major isoproteins each having the ability not only to bind [3H]-methotrexate but also reduce [3H]-pteroylglutamic acid to [3H]-tetrahydropteroylglutamic acid. This suggests that the two binders of methotrexate may be the two forms of dihydrofolate reductase having different affinities for this anticancer drug.
Iqbal
M P
MP
Department of Biochemistry, Aga Khan University, Karachi.
Mehboobali
N
N
Waqar
M A
MA
Ahmed
M
M
D002277:Carcinoma / Q000201:enzymology*; D003110:Colonic Neoplasms / Q000201:enzymology*; D003600:Cytosol / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008727:Methotrexate / Q000378:metabolism*; D013762:Tetrahydrofolate Dehydrogenase / Q000378:metabolism*; D014020:Tissue Extracts
D014020:Tissue Extracts; D013762:Tetrahydrofolate Dehydrogenase; D008727:Methotrexate