A fluorescence stopped-flow kinetic study of the conformational activation of α-chymotrypsin and several mutants

Protein Science

Volumn 13, Issue 9, 2004, Pages 2533-2540

  Verheyden, Gert ^a,b   Matrai, Janka ^a   Volckaert, Guido ^a   Engelborghs, Yves ^a  

^a UNIVERSITY OF LEUVEN   (Belgium)

^b INNOGENETICS   (Belgium)

Author keywords

Fluorescence stopped flow measurements; Structure function relationship; Targeted molecular dynamics; Tryptophan fluorescence

Indexed keywords

CHYMOTRYPSIN A; MUTANT PROTEIN; PROFLAVINE; TRYPTOPHAN;

ACTIVATION FREE ENERGY; ACTIVATION FREE ENTHALPY; ACTIVATION FREE ENTROPY CHANGE; ARTICLE; CONFORMATIONAL TRANSITION; ENERGY; ENTHALPY; ENTROPY; ENZYME ACTIVATION; ENZYME BINDING; ENZYME CONFORMATION; ENZYME KINETICS; FLUORESCENCE; FLUORESCENCE STOPPED FLOW MEASUREMENT; MEASUREMENT; MOLECULAR DYNAMICS; NONHUMAN; PARAMETER; PH; PRIORITY JOURNAL; STRUCTURE ACTIVITY RELATION; WILD TYPE;

ANIMALS; CATTLE; CHYMOTRYPSIN; ENZYME ACTIVATION; FLUORESCENCE; HYDROGEN-ION CONCENTRATION; KINETICS; MODELS, MOLECULAR; MUTATION; PROTEIN CONFORMATION; RATS; STRUCTURE-ACTIVITY RELATIONSHIP; TRYPTOPHAN;

BOVINAE;

EID: 4344569109     PISSN: 09618368     EISSN: None     Source Type: Journal    
DOI: 10.1110/ps.04709604     Document Type: Article

References (29)

1. Amneus, H., Gabel, D., and Kasche, V. 1976. Resolution in affinity chromatography. The effect of the heterogeneity of immobilized soybean trypsin inhibitor on the separation of pancreatic proteases. J. Chromatogr. 120: 391-397.
2. Aune, K.C., Goldsmith, L.C., and Timasheff, S.N. 1971. Dimerization of α-chymotrypsin. II. Ionic strength and temperature dependence. Biochemistry 10: 1617-1622.
3. Berendsen, H.J.C. and Hayward, S. 2000. Collective protein dynamics in relation to function. Curr. Opin. Struct. Biol. 10: 165-169.
4. Bevington, P.R. 1969. Data reduction and error analysis for the physical sciences. McGraw-Hill, New York.
5. Brothers, H.M. and Kostic, N.M. 1990. Catalytic activity of the serine proteases α-chymotrypsin and α-lytic protease tagged at the active-site with a (Terpyridine)Platinum(Ii) chromophore. Biochemistry 29: 7468-7474.
6. Chervenka, C.H. 1959. Ultraviolet spectral changes related to the enzymic activity of chymotrypsin. Biochim. Biophys. Acta 31: 85-95.
7. Delaage, M., Abita, J.P., and Lazdunski, M. 1968. Physico-chemical properties of bovine chymotrypsinogen B. A comparative study with trypsinogen and chymotrypsinogen A. Eur. J. Biochem. 5: 285-293.
8. Delmar, E.G., Largman, C., Brodrick, J.W., and Geokas, M.C. 1979. Sensitive new substrate for chymotrypsin. Anal. Biochem. 99: 316-320.
9. Desie, G., Boens, N., and Deschryver, F.C. 1986. Study of the time-resolved tryptophan fluorescence of crystalline α-chymotrypsin. Biochemistry 25: 8301-8308.
10. Dixon, G.H. and Neurath, H. 1957. Acylation of the enzymatic site of Δ-chymotrypsin by esters, acid anhydrides, and acid chlorides. J. Biol. Chem. 225: 1049-1059.
11. Fersht, A.R. and Requena, Y. 1971. Equilibrium and rate constants for the interconversion of two conformations of α-chymotrypsin. The existence of a catalytically inactive conformation at neutral pH. J. Mol. Biol 60: 279-290.
12. Hedstrom, L., Perona, J.J., and Rutter, W.J. 1994. Converting trypsin to chymotrypsin-Residue-172 is a substrate-specificity determinant. Biochemistry 33: 8757-8763.
13. Izrailev, S., Stepaniants, S., Isralewitz, B., Kosztin, D., Molnar, E., Lu, H., Wriggers, W., and Schulten, K. 1998. Steered molecular dynamics. In Computational molecular dynamics: Challenges, methods, ideas (eds. P. Deuflhard et al.), pp. 39-65. Springer Verlag, Berlin.
14. Kasche, V., Amneus, H., Gabel, D., and Naslund, L. 1977. Rapid zymogen activation and isolation of serine proteases from an individual mouse pancreas by affinity chromatography: Genetical heterogeneity of chymotrypsins of MMS musculus. Biochim. Biophys. Acta 490: 1-18.
15. Lambeir, A. and Engelborghs, Y. 1981. A fluorescence stopped flow study of colchicine binding to tubulin. J. Biol. Chem. 256: 3279-3282.
16. Peterman, B.F. 1979. Measurement of the dead time of a fluorescence stopped-flow instrument. Anal. Biochem. 93: 442-444.
17. Phillips, M.A., Fletterick, R., and Rutter, W.J. 1990. Arginine-127 stabilizes the transition-state in carboxypeptidase. J. Biol. Chem. 265: 20692-20698.
18. Sano, M., Tsukimura, T., and Yamazaki, A. 1988. Identification of acetylcholine, histamine and serotonin receptors in the porcine dental pulp. Showa. Shigakkai. Zasshi 8: 427-431.
19. Schlitter, J., Engels, M., Kruger, P., Jacoby, E., and Wollmer, A. 1993. Targeted molecular-dynamics simulation of conformational change-Application to the T[-]R transition in insulin. Mol. Simul. 10: 291-308.
20. Schlitter, J., Engels, M., and Kruger, P. 1994. Targeted molecular-dynamics-A new approach for searching pathways of conformational transitions. J. Mol. Graph. 12: 84-89.
21. Shearwin, K.E. and Winzor, D.J. 1990. Effect of calcium ion on the dimerization of α-chymotrypsin. Biochim. Biophys. Acta 1038: 136-138.
22. Steffen, L.W. and Steffen, B.W. 1976. Improved method for measuring fibrinogen in plasma, with use of a plasmin Inhibitor. Clin. Chem. 22: 381-383.
23. Stoesz, J.D. and Lumry, R.W. 1978. Refolding transition of α-chymotrypsin-pH and salt dependence. Biochemistry 17: 3693-3699.
24. Swegat, W., Krueger, P., and Schlitter, J. 1997. TMD-implementation in GROMOS96, pp. 1-11.
25. Venekei, I., Graf, L., and Rutter, W.J. 1996a. Expression of rat chymotrypsinogen in yeast: A study on the structural and functional significance of the chymotrypsinogen propeptide. FEBS Lett. 379: 139-142.
26. Venekei, I., Szilagyi, L., Graf, L., and Rutter, W.J. 1996b. Attempts to convert chymotrypsin to trypsin. FEBS Lett. 379: 143-147.
27. Verheyden, G., Volckaert, G., and Engelborghs, Y. 2000. Expression of chymotrypsin(ogen) in the thioredoxin reductase deficient mutant strain of Escherichia coli AD494(DE3) and purification via a fusion product with a hexahistidine-tail. J. Chromatogr. B 737: 213-224.
28. Wang, D.C., Bode, W., and Huber, R. 1985. Bovine chymotrypsinogen-A X-ray crystal-structure analysis and refinement of a new crystal form at 1.8 Å resolution. J. Mol. Biol. 185: 595-624.
29. Wroblowski, B., Diaz, J.F., Schlitter, J., and Engelborghs, Y. 1997. Modelling pathways of α-chymotrypsin activation and deactivation. Protein Eng. 10: 1163-1174.