Adjustment of conformational flexibility of glyceraldehyde-3-phosphate dehydrogenase as a means of thermal adaptation and allosteric regulation.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Thermotoga maritima (TmGAPDH) is a thermostable enzyme (Tm = 102 degrees C), which is fully active at temperatures near 80 degrees C but has very low activity at room temperature. In search for an explanation of this behavior, we measured the conformational flexibility of the protein by hydrogen-deuterium exchange and compared the results with those obtained with GAPDH from rabbit muscle (RmGAPDH). At room temperature, the conformational flexibility of TmGAPDH is much less than that of RmGAPDH, but increases with increasing temperature and becomes comparable to that of RmGAPDH near the physiological temperature of Thermotoga maritima. Using the available three-dimensional structures of the two enzymes, we compared the B factors that reflect the local mobility of protein atoms. The largest differences in B factors are seen in the coenzyme and NAD binding regions. The likely reason for the low activity of TmGAPDH at room temperature is that the motions required for enzyme functions are restricted. The findings support the idea of "corresponding states" which claims that over the time span of evolution, the overall conformational flexibility of proteins has been preserved at their corresponding physiological temperatures.
Hajdú
István
I
Institute of Enzymology, Biological Research Center, Hungarian Academy of Sciences, Karolina út 29-31, 1113, Budapest, Hungary.
Bothe
Csaba
C
Szilágyi
András
A
Kardos
József
J
Gál
Péter
P
Závodszky
Péter
P
D000222:Adaptation, Physiological; D000494:Allosteric Regulation; D000818:Animals; D041961:Deuterium Exchange Measurement; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D009132:Muscles / Q000201:enzymology; D011487:Protein Conformation; D011817:Rabbits; D013696:Temperature; D020124:Thermotoga maritima / Q000201:enzymology*
D005987:Glyceraldehyde-3-Phosphate Dehydrogenases