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Theory and practice of the analytical centrifugation of an active substrate–enzyme complex
Author(s) -
Cohen R.,
Giraud B.,
Messiah A.
Publication year - 1967
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.1967.360050208
Subject(s) - chemistry , substrate (aquarium) , diffusion , centrifugation , enzyme , sedimentation , enzyme assay , chromatography , analytical chemistry (journal) , thermodynamics , biochemistry , physics , paleontology , oceanography , biology , geology , sediment
The active enzyme centrifugal‐ion (AEC) method presented here permits the hydrodynamic study of active enzyme – substrate ( s ) complexes in solutions containing micro‐gram amounts of the studied enzyme, even if the enzyme preparation is very impure. The AEC method can he used only when the specific enzymatie reaction can be measured directly in a spectrophotometer. The general equations relevant to the method and their solutions are presented in detail. Their use requires some numerical calculations. A practical summary of the AEC method is given, and the precision of the measured values of the sedimentation and diffusion coefficients is discussed.