Thermodynamics and kinetics of the hydrolysis of the reactive-site peptide bond in pancreatic trypsin inhibitor (Kunitz) by Dermasterias imbricata trypsin 1

Biochemistry ◽  
1980 ◽  
Vol 19 (1) ◽  
pp. 131-137 ◽  
Author(s):  
David A. Estell ◽  
Karl A. Wilson ◽  
Michael Laskowski
Keyword(s):  
Trypsin Inhibitor ◽  
Peptide Bond ◽  
Reactive Site ◽  
Thermodynamics And Kinetics ◽  
Pancreatic Trypsin Inhibitor ◽  
Kinetics Of ◽  
Hydrolysis Of

scholarly journals Kinetics of tryptic hydrolysis of the arginine-valine bond in folded and unfolded ribonuclease T1

1984 ◽  
Vol 219 (2) ◽  
pp. 411-417 ◽  
Author(s):  
C N Pace ◽  
A J Barrett
Keyword(s):  
Kinetic Studies ◽  
Peptide Bond ◽  
Small Peptides ◽  
Ribonuclease T1 ◽  
Chemically Modified ◽  
Steady State Kinetic ◽  
Kinetics Of ◽  
Hydrolysis Of ◽  
State Kinetic ◽  
Tryptic Hydrolysis

We have used ribonuclease T1 and its chemically modified derivatives as substrates, and trypsin as proteinase, to investigate the kinetics of proteolysis of a specific peptide bond in the folded and unfolded conformations of a protein. Steady-state kinetic studies showed that Km = 0.27 mM and Kcat. = 2.45 s-1 for the tryptic hydrolysis of the Arg(77)-Val(78) peptide bond in unfolded ribonuclease T1. This Km is somewhat lower than, and the kcat. value similar to, values found for the tryptic hydrolysis of comparable bonds in small peptides. Our data for the initial velocity of hydrolysis of the Arg(77)-Val(78) bond in a solution of the folded protein indicate that the bond is at least 1700 times less rapidly hydrolysed in the folded than in the unfolded conformation of ribonuclease T1, and do not exclude the possibility that the bond is completely resistant to hydrolysis in the folded protein.

Stopped-flow kinetics of the resynthesis of the reactive site peptide bond in kallikrein inhibitor (Kunitz) by β-trypsin

Biochemistry ◽  
1978 ◽  
Vol 17 (9) ◽  
pp. 1675-1682 ◽  
Author(s):  
Ulrich Quast ◽  
Juergen Engel ◽  
Erna Steffen ◽  
Harald Tschesche ◽  
Sigrid Kupfer
Keyword(s):  
Peptide Bond ◽  
Stopped Flow ◽  
Reactive Site ◽  
Kallikrein Inhibitor ◽  
Stopped Flow Kinetics ◽  
Kinetics Of
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