19F-n.m.r. studies of 3',5'-difluoromethotrexate binding to Lactobacillus casei dihydrofolate reductase. Molecular motion and coenzyme-induced conformational changes
- PMID: 6424648
- PMCID: PMC1153266
- DOI: 10.1042/bj2170659
19F-n.m.r. studies of 3',5'-difluoromethotrexate binding to Lactobacillus casei dihydrofolate reductase. Molecular motion and coenzyme-induced conformational changes
Abstract
19F-n.m.r. spectroscopy was used to study the binding of 3',5'-difluoromethotrexate to dihydrofolate reductase (tetrahydrofolate dehydrogenase) from Lactobacillus casei. The benzoyl ring of the bound difluoromethotrexate was found to 'flip' about its symmetry axis, and the rate (7.3 X 10(3) s-1 at 298 K) and activation parameters for this process were determined by lineshape analysis of the 19F-n.m.r. spectrum at a series of temperatures in the range 273-308 K. The contributions to the barrier for this process are discussed. Addition of NADP+ or NADPH to form the enzyme-difluoromethotrexate-coenzyme ternary complex led to an increase in the rate of benzoyl ring flipping by a factor of 2.6-2.8-fold, and to substantial changes in the 19F-n.m.r. chemical shifts. The possible nature of the coenzyme-induced conformational changes responsible for these effects is discussed.
Similar articles
-
A 1H n.m.r. study of the role of the glutamate moiety in the binding of methotrexate to Lactobacillus casei dihydrofolate reductase.Br J Pharmacol. 1984 Feb;81(2):309-15. doi: 10.1111/j.1476-5381.1984.tb10080.x. Br J Pharmacol. 1984. PMID: 6423020 Free PMC article.
-
Structural comparisons of complexes of methotrexate analogues with Lactobacillus casei dihydrofolate reductase by two-dimensional 1H NMR at 500 MHz.Biochemistry. 1987 Dec 29;26(26):8585-90. doi: 10.1021/bi00400a014. Biochemistry. 1987. PMID: 3126805
-
Multinuclear NMR characterization of two coexisting conformational states of the Lactobacillus casei dihydrofolate reductase-trimethoprim-NADP+ complex.Biochemistry. 1984 Sep 25;23(20):4733-42. doi: 10.1021/bi00315a032. Biochemistry. 1984. PMID: 6437442
-
Folic acid and vitamin B12: transport and conversion to coenzyme forms.Adv Enzyme Regul. 1974;12:131-53. doi: 10.1016/0065-2571(74)90011-9. Adv Enzyme Regul. 1974. PMID: 4156820 Review. No abstract available.
-
Dihydrofolate reductases: structural and mechanistic aspects.Ann N Y Acad Sci. 1971 Nov 30;186:85-99. Ann N Y Acad Sci. 1971. PMID: 4400072 Review. No abstract available.
Cited by
-
3H-n.m.r. studies of multiple conformations and dynamic processes in complexes of folate and methotrexate with Lactobacillus casei dihydrofolate reductase.Biochem J. 1994 Oct 15;303 ( Pt 2)(Pt 2):401-5. doi: 10.1042/bj3030401. Biochem J. 1994. PMID: 7980397 Free PMC article.
-
Structure and dynamics in solution of the complex of Lactobacillus casei dihydrofolate reductase with the new lipophilic antifolate drug trimetrexate.Protein Sci. 1999 Mar;8(3):467-81. doi: 10.1110/ps.8.3.467. Protein Sci. 1999. PMID: 10091649 Free PMC article.
-
The study of conformational states of proteins by nuclear magnetic resonance.Biochem J. 1985 Oct 1;231(1):1-10. doi: 10.1042/bj2310001. Biochem J. 1985. PMID: 2998335 Free PMC article. Review.
-
19F Magic Angle Spinning NMR Spectroscopy and Density Functional Theory Calculations of Fluorosubstituted Tryptophans: Integrating Experiment and Theory for Accurate Determination of Chemical Shift Tensors.J Phys Chem B. 2018 Jun 14;122(23):6148-6155. doi: 10.1021/acs.jpcb.8b00377. Epub 2018 May 30. J Phys Chem B. 2018. PMID: 29756776 Free PMC article.
-
Dynamics of trimethoprim bound to dihydrofolate reductase.Proc Natl Acad Sci U S A. 1988 Jun;85(11):3787-91. doi: 10.1073/pnas.85.11.3787. Proc Natl Acad Sci U S A. 1988. PMID: 3131763 Free PMC article.
References
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
