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Theory and practice of the analytical centrifugation of an active substrate–enzyme complex

✍ Scribed by R. Cohen; B. Giraud; A. Messiah

Publisher
Wiley (John Wiley & Sons)
Year
1967
Tongue
English
Weight
1008 KB
Volume
5
Category
Article
ISSN
0006-3525

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✦ Synopsis

The active eiizyme centrifugat.ion (AEC) method preseiited here permits the hydrodynamic study of active enzyme-substrate(s) complexes in solutions containing microgram am0unt.s of the studied enzyme, even if the enzyme preparation is very impure. The AEC method can be used only when the specific enzymatic reaction can be measured directly in a spectrophotometer. The general equations relevant. to the method and their solutions are presented in detail.

A practical summary of the AEC method is given, and the precisioii of the measured valries of t,he sedimentation and diffusion coefficients is discussed. Their use requires some numerical calculatioiis.

* An error in the baselinevalue changes the results for N1'(mar) and p'in the same direction. Since 2' depends only on their ratio we improved the precision by using for the calculation of each Z'the experimentally measured N I ' ( B Y ) and the experimentally determined p' (and not the least-square average p ' ) . Figure 5 shows clearly that the deviations are smaller than the 10% mentioned above.

📜 SIMILAR VOLUMES

Sedimentation of generalized systems of
Sedimentation of generalized systems of interacting particles. II. Active enzyme centrifugation—theory and extensions of its validity range
✍ René Cohen; Jean-Michel Claverie 📂 Article 📅 1975 🏛 Wiley (John Wiley & Sons) 🌐 English ⚖ 837 KB

## Abstract The first detailed application of a recently published very general approach to chemical equilibria during sedimentation is presented. As a consequence of the very extensive theoretical treatment, made possible by this approach, the active enzyme analytical centrifugation method can now