Conversion of trypsin into a Na(+)-activated enzyme

Biochemistry
Michael J PageEnrico Di Cera

Abstract

Serine proteases of the chymotrypsin family show a dichotomous amino acid distribution for residue 225. Enzymes carrying Tyr at position 225 are activated by Na(+), whereas those carrying Pro are devoid of Na(+) binding and activation. Previous studies have demonstrated that the Y225P conversion is sufficient to abrogate Na(+) activation in several enzymes. However, the reverse substitution P225Y is necessary but not sufficient to introduce Na(+) binding and activation. Here we report that Streptomyces griseus trypsin, carrying Pro-225, can be engineered into a Na(+)-activated enzyme by replacing residues in the 170, 186, and 220 loops to those of coagulation factor Xa. The findings represent the first instance of an engineered Na(+)-activated enzyme and a proof of principle that should enable the design of other proteases with enhanced catalytic activity and allosteric regulation mediated by monovalent cation binding.

References

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Nov 4, 2005·The Journal of Biological Chemistry·Enrico Di Cera

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Citations

Dec 12, 2012·Quarterly Reviews of Biophysics·Pengyu RenNathan A Baker
Mar 10, 2007·Physical Chemistry Chemical Physics : PCCP·Enrico Di CeraLaura C Garvey
Feb 11, 2015·Biochemistry·Leslie A PelcEnrico Di Cera
Apr 20, 2010·Journal of Molecular Biology·Michael J Page, Enrico Di Cera
Oct 4, 2006·Physiological Reviews·Michael J Page, Enrico Di Cera
Apr 13, 2007·The Journal of Biological Chemistry·Francesca MarinoEnrico Di Cera
Jul 20, 2007·The Journal of Biological Chemistry·Leslie A Bush-PelcEnrico Di Cera
Jan 25, 2014·The Journal of Biological Chemistry·Shilpa GadwalMaria Sandkvist

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