Minimal functional unit of lactate dehydrogenase.
The tetrameric heart isozyme of lactate dehydrogenase (H4) is modified by p-chloromercuribenzoate (PCMB) to produce the inactive tetramer (H4) and then hybridized with native tetrameric muscle isozyme (M4). The hybrid mixture (M4, H'M3, H2'M2, H3'M, and H4') was isolated by polyacrylamide gel electrophoresis (PAGE) and then stained for enzyme activity and with Coomassie brilliant blue. Only three bands were found on the gels in either case. The hybrid enzymes (H'M3 and H2'M2) as isolated by PAGE have half the specific activity of the native muscle enzyme. The electrophoresis properties of H'M3 are very similar to those of HM3, while the electrophoresis properties of H2'M2 are very similar to those of H2M2. The above results strongly suggest that the tetramer having enzymatic activity contains at least two native subunits, and the di-subunit in the tetrameric enzyme is the minimal functional unit.
Wang
X C
XC
Department of Biological Science and Biotechnology, School of Life Science and Engineering, Tsinghua University, Beijing, China.
Jiang
L
L
Zhou
H M
HM
D000818:Animals; D002729:Chloromercuribenzoates; D004591:Electrophoresis, Polyacrylamide Gel; D007527:Isoenzymes; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D046911:Macromolecular Substances; D008961:Models, Structural; D018482:Muscle, Skeletal / Q000737:chemistry; D009206:Myocardium / Q000201:enzymology; D055503:Protein Multimerization; D013552:Swine; D020245:p-Chloromercuribenzoic Acid
D002729:Chloromercuribenzoates; D007527:Isoenzymes; D046911:Macromolecular Substances; D020245:p-Chloromercuribenzoic Acid; D007770:L-Lactate Dehydrogenase