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Studies of Amino-Acid Sequence in Dihydrofolate Reductase from a Human Methotrexate-Resistant Cell Line KB/6b.
- Source :
-
European Journal of Biochemistry . 5/2/83, Vol. 132 Issue 2, p351-359. 9p. - Publication Year :
- 1983
-
Abstract
- The partial ammo acid sequence of dihydrofolate reductase (DHFR. EC 1.5,1.3) from human KB/6b cells hi been determined by using 3.5 mg of protein. Peptides covering the entire polypeptide chain were recovered from preparative peptide maps generated by the combination of paper chromatography and electrophoresis at pH 4.4 Peptide maps from mouse L1210 DHFR were also generated for comparison. Amino acid sequence of 75% of the 186 amino acid residues in the polypeptide chain of human KB/6b DHFR was obtained from Edman degradation and the remaining sequence was deduced from the amino acid compositions, from electrophoretic mobilities of related peptides and from the sequence homologies with other known mammalian DHFR sequence. A comparison of the proposed human DHFR sequence with the previously known sequent of mouse enzyme [Stone, et al. (1979) J. Biol. Chem. 245, 480-488] indicates that 18 differences are located in the established sequence of 139 residues and that 5 additional differences are in the tentative sequence of the remaining 47 amino acids. Kinetic properties of human KB/6b and mouse L1210 DHFR, which were determined in parallel experiments, are also compared. The possible structural-functional relationships between human and mouse DHFR are discussed. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 00142956
- Volume :
- 132
- Issue :
- 2
- Database :
- Academic Search Index
- Journal :
- European Journal of Biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 15817938
- Full Text :
- https://doi.org/10.1111/j.1432-1033.1983.tb07369.x