The studies of cooperative regions in T7 RNA polymerase.
The heat denaturation of bacteriophage T7 RNA polymerase (T7RNAP) was studied by scanning microcalorimetry. The thermodynamic parameters of the denaturation were estimated within the pH range 6-9. The analysis of the denaturation curves showed the presence of two cooperative parts of the T7RNAP molecule melting according to the 'all-or-none' principle. The molecular masses of these parts were determined as 22 and 77 kDa. These values are close to the molecular masses of protein domains obtained from X-ray diffraction and limited trypsinolysis data. The smaller N-terminal domain was shown to increase the thermostability of the 'catalytic' C-terminal domain within the intact T7RNAP molecule.
Protasevich
I I
II
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow.
Memelova
L V
LV
Kochetkov
S N
SN
Makarov
A A
AA
D017123:Bacteriophage T7 / Q000201:enzymology; D002152:Calorimetry, Differential Scanning; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry*; D011487:Protein Conformation; D011489:Protein Denaturation; D013816:Thermodynamics; D014764:Viral Proteins
D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases