Abstract
Six mutations determining resistance to amethopterin were examined for their effects on the active transport of the drug. In strains bearing each of the mutations and exhibiting resistance levels varying from 10- to 100-fold, transport at limiting concentrations of H3-amethopterin was reduced from 2.5 to 10 times the rate characteristic of the wild type. Kinetic analysis of transport showed an increase in the value for Km of the system in all of the mutants. Values for the wild-type system were 0.9 × 10−6m and for the mutants varied between 2.5 × 10−6m and 9.0 × 10−6m. Values for Vmax were approximately the same for each system. The mutant transport systems also exhibited a shift in pH optimum from near 6.0 (wild-type) to below 5.0. The results were interpreted as an alteration in the binding properties of the permease in the mutant strains.
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Selected References
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- COHEN G. N., MONOD J. Bacterial permeases. Bacteriol Rev. 1957 Sep;21(3):169–194. doi: 10.1128/br.21.3.169-194.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ephrussi-Taylor H., Sicard A. M., Kamen R. Genetic Recombination in DNA-Induced Transformation of Pneumococcus. I. the Problem of Relative Efficiency of Transforming Factors. Genetics. 1965 Mar;51(3):455–475. doi: 10.1093/genetics/51.3.455. [DOI] [PMC free article] [PubMed] [Google Scholar]
- FISCHER G. A. Detective transport of amethopterin (methotrexate) as a mechanism of resistance to the antimetabolite in L5178Y leukemic cells. Biochem Pharmacol. 1962 Dec;11:1233–1234. doi: 10.1016/0006-2952(62)90200-9. [DOI] [PubMed] [Google Scholar]
- KOCH A. L. THE ROLE OF PERMEASE IN TRANSPORT. Biochim Biophys Acta. 1964 Jan 27;79:177–200. doi: 10.1016/0926-6577(64)90050-6. [DOI] [PubMed] [Google Scholar]
- Kessel D., Hall T. C., Roberts D., Wodinsky I. Uptake as a determinant of methotrexate response in mouse leukemias. Science. 1965 Nov 5;150(3697):752–754. doi: 10.1126/science.150.3697.752. [DOI] [PubMed] [Google Scholar]
- PINE M. J. Uptake of aminopterin by Bacillus subtilis. J Bacteriol. 1960 Jun;79:827–834. doi: 10.1128/jb.79.6.827-834.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SIROTNAK F. M., DONATI G. J., HUTCHISON D. J. DIHYDROFOLATE REDUCTASE IN GENOTYPICALLY DISTINGUISHABLE AMETHOPTERIN-RESISTANT DIPLOCOCCUS PNEUMONIAE. J Biol Chem. 1964 Aug;239:2677–2682. [PubMed] [Google Scholar]
- SIROTNAK F. M., DONATI G. J., HUTCHISON D. J. GENETIC MODIFICATION OF THE STRUCTURE AND AMOUNT OF DIHYDROFOLATE REDUCTASE IN AMETHOPTERIN-RESISTANT DIPLOCOCCUS PNEUMONIAE. J Biol Chem. 1964 Dec;239:4298–4302. [PubMed] [Google Scholar]
- SIROTNAK F. M., LUNT R. B., HUTCHISON D. J. THE DISTRIBUTION OF MUTATIONAL SITES AFFECTING RESISTANCE TO AMETHOPTERIN IN DIPLOCOCCUS PNEUMONIAE. Genetics. 1964 Mar;49:439–452. doi: 10.1093/genetics/49.3.439. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SIROTNAK F. M., LUNT R. B., HUTCHISON D. J. Transformation of resistance to 8-azaguanine in diplococcus pneumoniae. Nature. 1960 Aug 27;187:800–801. doi: 10.1038/187800a0. [DOI] [PubMed] [Google Scholar]
- Sicard A. M., Ephrussi-Taylor H. Genetic recombination in DNA-induced transformation of Pneumococcus. II. Mapping the amiA region. Genetics. 1965 Dec;52(6):1207–1227. doi: 10.1093/genetics/52.6.1207. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sirotnak F. M., Hutchison D. J. Purification and properties of mutant and wild-type dihydrofolate reductase from Diplococcus pneumoniae. J Biol Chem. 1966 Jun 25;241(12):2900–2906. [PubMed] [Google Scholar]
- Sirotnak F. M., Sargent M. G., Hutchison D. J. Genetically alterable transport of amethopterin in Diplococcus pneumoniae. I. Physiological properties and kinetics of the wild-type system. J Bacteriol. 1967 Jan;93(1):309–314. doi: 10.1128/jb.93.1.309-314.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WERKHEISER W. C. THE BIOCHEMICAL, CELLULAR, AND PHARMACOLOGICAL ACTION AND EFFECTS OF THE FOLIC ACID ANTAGONISTS. Cancer Res. 1963 Sep;23:1277–1285. [PubMed] [Google Scholar]
