Biochemistry of Vitamin B6 pp 183-186 | Cite as
Phosphorus-31 NMR Study of Pyridoxal 5’-Phosphate Binding to Escherichia Coli Tryptophan Synthase Modified by Limited Proteolysis
Summary
The β2 subunit of α2β2 tryptophan synthase from Escherichia coli has been proteolyzed by limited treatment with protease from Staphylococcus aureus, V8. Interaction of this derivative (nicked β2) with pyridoxal-P has been investigated using 31P nuclear magnetic resonance (NMR). The phosphate signal of pyridoxal-P shows a linewidth significantly smaller than expected for a rigidly bound cofactor molecule. Upon addition of the corresponding a subunit further line narrowing is observed indicating that the nicked β2 protein is still capable to form a native-like α2β2 complex. These results are compared with data for both the native β2 subunit and the α2 holo β2 complex.
Keywords
Nuclear Magnetic Resonance Phosphate Binding Limited Proteolysis Physiological Chemistry Cleave Peptide BondPreview
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References
- Ahmed S., Fairwell T., Dunn S., Kirschner K., and Miles E.W. (1986) Biochemistry 25, 3118–3124CrossRefGoogle Scholar
- Bartholmes, P., Kirschner, K., and Gschwind, H.-P. (1976) Biochemistry 15, 4712–4717CrossRefGoogle Scholar
- Faeder, E.J., and Hammes, G.G. (1970) Biochemistry 9, 4043–4049CrossRefGoogle Scholar
- Högberg-Raibaud, A., and Goldberg, M.E. (1978) Proc. Natl. Acad. Sci. USA 74, 442–446CrossRefGoogle Scholar
- Kirschner, K., Wiskocil, R., Foehn, M., and Rezeau, L. (1975) Eur. J. Biochem. 60, 513–523CrossRefGoogle Scholar
- Schnackerz, K.D., and Bartholmes, P. (1983) Biochem. Biophys. Res. Comm. 111, 817–823CrossRefGoogle Scholar
- Tate, K.M., Higgins, D.L., Holmes, W.E., Winkler, M.E., Heyneker, H.L., and Vehar, G.A. (1987) Biochemistry 26, 338–343CrossRefGoogle Scholar