Open AccessNew Protein Reagents. N ‐(4‐Chloromercuriphenyl)‐4‐Chloro‐3,5‐Dinitrobenzamide and Its Use as a Probe of the Quaternary Structure of Yeast Alcohol Dehydrogenase
Author(s) -
DIOPOH Jacques,
OLOMUCKI Martin
Publication year - 1977
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1977.tb11546.x
Subject(s) - protein quaternary structure , dimer , reagent , chemistry , bifunctional , monomer , moiety , alcohol dehydrogenase , dehydrogenase , enzyme , yeast , stereochemistry , alcohol , biochemistry , organic chemistry , polymer , catalysis , protein subunit , gene
The new bifunctional reagent, N ‐(4‐chloromercuriphenyl)‐4‐chloro‐3,5‐dinitrobenzamide (I) was used to investigate the quaternary structure of yeast alcohol dehydrogenase. The four essential – SH groups of the enzyme were substituted by the mercuriphenyl moiety of compound I in the course of the reaction of one mole of protein with four moles of the reagent (one molecule of compound I incorporated by yeast alcohol dehydrogenase monomer). In a second step only two of the four chlorodinitrophenyl fragments bound to the protein established intermonomeric cross‐links with non‐essential – NH 2 groups. The resulting dimers could be re‐dissociated with mercaptoethanol. This result suggests that the four protomers of the enzyme could be arranged as a dimer of dimers.