The role of bound calcium ions in thermostable, proteolytic enzymes. II. Studies on thermolysin, the thermostable protease from Bacillus thermoproteolyticus.

Biochemistry
G Voordouw, R S Roche

Abstract

The functional properties of the four calcium ions, bound by thermolysin, appear to be very similar to those of the single calcium ion bound by thermomycolase (G. Voordouw and R.S. Roche (1975), Biochemistry, preceding paper in this issue). Hence when the free calcium ion concentration is varied in the range where the calcium double-site dissociates (G. Voordouw and R.S. Roche (1974), Biochemistry 13, 5017), no changes are observed in the sedimentation coefficient or the peptide circular dichroism. Differences in molar ellipticity and molar extinction coefficient occur in the aromatic ultraviolet region, which parallel the occupancy of the calcium binding double site. The difference spectrum, characterized by a main band at 290 nm and a somewhat smaller band at 283 nm, is interpreted as due to the transfer of a partially buried tryptophan residue to the aqueous solvent upon dissociation of the two calcium ions from the double site. This is most likely Trp-186, which is in between Asp-185 and Glu-187, two chelating amino acids of this site. From the calcium dependence of the rate constant for autolytic degradation we conclude, as for thermomycolase, that only conformers devoid of bound calcium ion serve as substrates in the reac...Continue Reading

Citations

Aug 1, 1993·Journal of Computer-aided Molecular Design·G Vriend, V Eijsink
Aug 24, 2004·Extremophiles : Life Under Extreme Conditions·Xiao-Ge ChenStephen Tiong-Lee Tay
Oct 15, 1987·Biochimica Et Biophysica Acta·Z Wasylewski, M R Eftink
Mar 1, 1986·Biophysical Chemistry·J D BuchananR S Roche
Oct 23, 2012·Journal of Biochemistry·Yuichi KawasakiKuniyo Inouye
Jan 20, 2010·Protein Engineering, Design & Selection : PEDS·Hiroyasu OginoNoriyuki Doukyu
Jun 2, 2010·Protein Engineering, Design & Selection : PEDS·S Mohsen AsghariKhosro Khajeh
Sep 15, 1988·European Journal of Biochemistry·J BajorathW Saenger
Jan 1, 1985·Annals of the New York Academy of Sciences·D G HangauerG M Smith
Jan 1, 1983·Biopolymers·R J Corbett, R S Roche
Jul 1, 1980·Archives of Biochemistry and Biophysics·R A SmithP H Kopper
Oct 15, 1992·Biochemical and Biophysical Research Communications·J S JangS M Byun
Jan 1, 1978·CRC Critical Reviews in Microbiology·R E Amelunxen, A L Murdock
Jan 1, 1978·CRC Critical Reviews in Biochemistry·R S Roche, G Voordouw

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