Key role of barstar Cys-40 residue in the mechanism of heat denaturation of bacterial ribonuclease complexes with barstar.
The mechanism by which barnase and binase are stabilized in their complexes with barstar and the role of the Cys-40 residue of barstar in that stabilization have been investigated by scanning microcalorimetry. Melting of ribonuclease complexes with barstar and its Cys-82-Ala mutant is described by two 2-state transitions. The lower-temperature one corresponds to barstar denaturation and the higher-temperature transition to ribonuclease melting. The barstar mutation Cys-40-Ala, which is within the principal barnase-binding region of barstar, simplifies the melting to a single 2-state transition. The presence of residue Cys-40 in barstar results in additional stabilization of ribonuclease in the complex.
Protasevich
I I
II
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow.
Schulga
A A
AA
Vasilieva
L I
LI
Polyakov
K M
KM
Lobachov
V M
VM
Hartley
R W
RW
Kirpichnikov
M P
MP
Makarov
A A
AA
D000409:Alanine / Q000235:genetics; D001407:Bacillus / Q000201:enzymology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D003545:Cysteine / Q000235:genetics / Q000378:metabolism*; D004722:Endoribonucleases / Q000378:metabolism*; D004795:Enzyme Stability; D006361:Heating; D016297:Mutagenesis, Site-Directed; D011489:Protein Denaturation; D012260:Ribonucleases / Q000378:metabolism*
D001426:Bacterial Proteins; C057438:barstar protein, Bacillus amyloliquefaciens; D004722:Endoribonucleases; D012260:Ribonucleases; C019760:Bacillus amyloliquefaciens ribonuclease; C022543:ribonuclease T(2); D003545:Cysteine; D000409:Alanine