Summary
Pyridoxal kinase (PK) catalyzes the formation of pyridoxal-5-phosphate (PLP) from pyridoxal (PL), ATP and a divalent cation (Zn2+). So far, there is no three-dimensional structure of PK available. Site-directed mutagenesis was carried out to study the importance of three conserved residues: Tyr137, Gly242 and G1y244. The mutants (Y137F, G242A and G244A) were constructed, expressed, purified and analyzed. The results demonstrated that the mutants had much less activity but with no dramatic variation in protein stability. Tyrl 37 residue was shown to be involved in PL binding but not ATP binding. For the G244A mutant, the absence of enzyme activity was probably due to the deficiency in PL binding rather than the lack of ATP binding. In the case of the G242A mutant, it did not bind to ATP or PL.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsPreview
Unable to display preview. Download preview PDF.
References
McCormick, D.B. and Snell, E. E. (1959).Proc. Natl. Acad. Sci. USA45, 1371–1379.
Kerry, J. A., Rohde, M. and Kwok, F. (1986).Eur. J. Biochem.158, 581–585.
Gao, Z.G., Lau, C. K., Lo, S. C. L., Choi, S. Y., Churchich, J. E. and Kwok, F. (1998).Int. J. Biochem. & Cell. Bio.30, 1379–1388.
Bradford, M. M. (1976).Anal. Biochem.72, 248–254.
Walker, J. E., Saraste, M., Runswick, M. J.and Gay, N. (1982).EMBOJ.1, 945–951.
Traut, T. W. (1994).Eur. J. Biochem.222, 9–19.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2000 Springer Basel AG
About this paper
Cite this paper
Leung, Y.C., Wong, H.Y., Churchich, J.E., Lo, S.C.L., Kwok, F. (2000). Structure-Function Relationships of Porcine Pyridoxal Kinase. In: Iriarte, A., Martinez-Carrion, M., Kagan, H.M. (eds) Biochemistry and Molecular Biology of Vitamin B6 and PQQ-dependent Proteins. Birkhäuser, Basel. https://doi.org/10.1007/978-3-0348-8397-9_45
Download citation
DOI: https://doi.org/10.1007/978-3-0348-8397-9_45
Publisher Name: Birkhäuser, Basel
Print ISBN: 978-3-0348-9549-1
Online ISBN: 978-3-0348-8397-9
eBook Packages: Springer Book Archive