Direct kinetics of bait region cleavage of α‐2‐macroglobulin by a rapid quenching method
- 1 July 1993
- journal article
- Published by Wiley in FEBS Letters
- Vol. 326 (1-3) , 80-82
- https://doi.org/10.1016/0014-5793(93)81765-r
Abstract
The rate of bait region cleavage of human α-2-macroglobulin by chymotrypsin was determined by a rapid quenching method under conditions where the bimolecular encounter between the two reactants was not rate-limiting. α2M was first mixed with a 30 molar excess of chymotrypsin in a sequential stopped-flow apparatus and after programmed time intervals the activity of chymotrypsin was quenched with 1 N HCl. The fraction of uncleaved subunits was quantitated by SDS-PAGE under reducing conditions. The result indicated that the bait region cleavage proceeded following a two-exponential decay curve with respective rate constants of k 1 = 40 s−1 and k 2 = 2 s−1.Keywords
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