Antibodies specific to modified glyceraldehyde-3-phosphate dehydrogenase induce inactivation of the native enzyme and change its conformation.
The antibodies specific to an inactive glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Bacillus stearothermophilus prepared by the treatment of the tetrameric holoenzyme with glutaraldehyde were obtained. They were purified from the pool of polyclonal rabbit antibodies to GAPDH with the use of immobilized GAPDH cross-linked by glutaraldehyde as an affinity sorbent. Such antibodies were capable of interacting with the native enzyme, inducing its time-dependent inactivation; the effect was different with the apo- and holoenzyme forms. Differential scanning calorimetry of the purified [GAPDH].[antibody] complex revealed a large shift of the temperature corresponding to the maximal heat capacity of the holoenzyme towards the lower temperature. Again, the effect appeared to be different with the apoenzyme. Together, the results are consistent with the hypothesis that a specific antibody is able to exercise a certain strain on the target protein, altering its conformation toward the structure of the species which served to select the antibody. The possibility of preparing selective enzyme inhibitors based on the antibodies specific to inactive enzyme conformations is considered.
Arutyunova
Elena I
EI
A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Lenin's Hills, Moscow 19992, Russia.
Arutyunov
Denis Yu
DY
Pleten'
Anatoliy P
AP
Nagradova
Natalya K
NK
Muronetz
Vladimir I
VI
D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000494:pharmacology*; D002152:Calorimetry, Differential Scanning; D001411:Geobacillus stearothermophilus / Q000201:enzymology*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D011487:Protein Conformation / Q000187:drug effects; D017510:Protein Folding; D020673:Protein Renaturation; D011817:Rabbits; D013696:Temperature
D000906:Antibodies; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases