The Use of Natural and Unnatural Amino Acid Substrates to Define the Substrate Specificity Differences of Escherichia Coli Aspartate and Tyrosine Aminotransferases

Overview

Journal Protein Sci

Specialty Biochemistry

Date 1995 Sep 1

PMID 8528072

Citations 5

Authors

J J Onuffer

B T Ton

I Klement

J F KIRSCH

Affiliations

Soon will be listed here.

Abstract

The tyrosine (eTATase) and aspartate (eAATase) aminotransferases of Escherichia coli transaminate diacarboxylic amino acids with similar rate constants. However, eTATase exhibits approximately 10(2)-10(4)-fold higher second-order rate constants for the transamination of aromatic amino acids than does eAATase. A series of natural and unnatural amino acid substrates was used to quantitate specificity differences for these two highly related enzymes. A general trend toward lower transamination activity with increasing side-chain length (extending from aspartate to glutamate to alpha-aminoadipate) is observed for both enzymes. This result suggests that dicarboxylate ligands associate with the two highly related enzymes in a similar manner. The high reactivity of the enzymes with L-Asp and L-Glu can be attributed to an ion pair interaction between the side-chain carboxylate of the amino acid substrate and the guanidino group of the active site residue Arg 292 that is common to both enzymes. A strong linear correlation between side-chain hydrophobicity and transamination rate constants obtains for n-alkyl side-chain amino substrates with eTATase, but not for eAATase. The present kinetic data support a model in which eAATase contains one binding mode for all classes of substrate, whereas the active site of eTATase allows an additional mode that has increased affinity for hydrophobic amino acid.

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References

Mullis K, Faloona F . Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987; 155:335-50. DOI: 10.1016/0076-6879(87)55023-6. View

MAVRIDES C . Transamination of aromatic amino acids in Escherichia coli. Methods Enzymol. 1987; 142:253-67. DOI: 10.1016/s0076-6879(87)42035-1. View

Hayashi H, Kuramitsu S, Inoue Y, Morino Y, KAGAMIYAMA H . [Arg292----Val] or [Arg292----Leu] mutation enhances the reactivity of Escherichia coli aspartate aminotransferase with aromatic amino acids. Biochem Biophys Res Commun. 1989; 159(1):337-42. DOI: 10.1016/0006-291x(89)92443-1. View

Seville M, Vincent M, Hahn K . Modeling the three-dimensional structures of bacterial aminotransferases. Biochemistry. 1988; 27(22):8344-9. DOI: 10.1021/bi00422a009. View

Julin D, KIRSCH J . Kinetic isotope effect studies on aspartate aminotransferase: evidence for a concerted 1,3 prototropic shift mechanism for the cytoplasmic isozyme and L-aspartate and dichotomy in mechanism. Biochemistry. 1989; 28(9):3825-33. DOI: 10.1021/bi00435a031. View

Mehta P, Hale T, Christen P . Evolutionary relationships among aminotransferases. Tyrosine aminotransferase, histidinol-phosphate aminotransferase, and aspartate aminotransferase are homologous proteins. Eur J Biochem. 1989; 186(1-2):249-53. DOI: 10.1111/j.1432-1033.1989.tb15202.x. View

Hayashi H, Inoue Y, Kuramitsu S, Morino Y, KAGAMIYAMA H . Effects of replacement of tryptophan-140 by phenylalanine or glycine on the function of Escherichia coli aspartate aminotransferase. Biochem Biophys Res Commun. 1990; 167(2):407-12. DOI: 10.1016/0006-291x(90)92037-z. View

Herold M, Kirschner K . Reversible dissociation and unfolding of aspartate aminotransferase from Escherichia coli: characterization of a monomeric intermediate. Biochemistry. 1990; 29(7):1907-13. DOI: 10.1021/bi00459a035. View

Kuramitsu S, Hiromi K, Hayashi H, Morino Y, KAGAMIYAMA H . Pre-steady-state kinetics of Escherichia coli aspartate aminotransferase catalyzed reactions and thermodynamic aspects of its substrate specificity. Biochemistry. 1990; 29(23):5469-76. DOI: 10.1021/bi00475a010. View

10.

Kamitori S, Okamoto A, Hirotsu K, Higuchi T, Kuramitsu S, KAGAMIYAMA H . Three-dimensional structures of aspartate aminotransferase from Escherichia coli and its mutant enzyme at 2.5 A resolution. J Biochem. 1990; 108(2):175-84. DOI: 10.1093/oxfordjournals.jbchem.a123178. View

11.

Danishefsky A, Onnufer J, Petsko G, Ringe D . Activity and structure of the active-site mutants R386Y and R386F of Escherichia coli aspartate aminotransferase. Biochemistry. 1991; 30(7):1980-5. DOI: 10.1021/bi00221a035. View

12.

Jager J, Solmajer T, Jansonius J . Computational approach towards the three-dimensional structure of E. coli tyrosine aminotransferase. FEBS Lett. 1992; 306(2-3):234-8. DOI: 10.1016/0014-5793(92)81007-9. View

13.

Toney M, KIRSCH J . Brønsted analysis of aspartate aminotransferase via exogenous catalysis of reactions of an inactive mutant. Protein Sci. 1992; 1(1):107-19. PMC: 2142083. DOI: 10.1002/pro.5560010111. View

14.

Hayashi H, Inoue K, Nagata T, Kuramitsu S, KAGAMIYAMA H . Escherichia coli aromatic amino acid aminotransferase: characterization and comparison with aspartate aminotransferase. Biochemistry. 1993; 32(45):12229-39. DOI: 10.1021/bi00096a036. View

15.

Malashkevich V, Toney M, Jansonius J . Crystal structures of true enzymatic reaction intermediates: aspartate and glutamate ketimines in aspartate aminotransferase. Biochemistry. 1993; 32(49):13451-62. DOI: 10.1021/bi00212a010. View

16.

Onuffer J, KIRSCH J . Characterization of the apparent negative co-operativity induced in Escherichia coli aspartate aminotransferase by the replacement of Asp222 with alanine. Evidence for an extremely slow conformational change. Protein Eng. 1994; 7(3):413-24. DOI: 10.1093/protein/7.3.413. View

17.

Jager J, Moser M, Sauder U, Jansonius J . Crystal structures of Escherichia coli aspartate aminotransferase in two conformations. Comparison of an unliganded open and two liganded closed forms. J Mol Biol. 1994; 239(2):285-305. DOI: 10.1006/jmbi.1994.1368. View

18.

Okamoto A, Higuchi T, Hirotsu K, Kuramitsu S, KAGAMIYAMA H . X-ray crystallographic study of pyridoxal 5'-phosphate-type aspartate aminotransferases from Escherichia coli in open and closed form. J Biochem. 1994; 116(1):95-107. DOI: 10.1093/oxfordjournals.jbchem.a124509. View

19.

Malashkevich V, Onuffer J, KIRSCH J, Jansonius J . Alternating arginine-modulated substrate specificity in an engineered tyrosine aminotransferase. Nat Struct Biol. 1995; 2(7):548-53. DOI: 10.1038/nsb0795-548. View

20.

Onuffer J, KIRSCH J . Redesign of the substrate specificity of Escherichia coli aspartate aminotransferase to that of Escherichia coli tyrosine aminotransferase by homology modeling and site-directed mutagenesis. Protein Sci. 1995; 4(9):1750-7. PMC: 2143225. DOI: 10.1002/pro.5560040910. View