Conformational transition of restriction endonuclease MvaI-substrate complex under the influence of Mg2+ probed by DNA-protein cross-linking studies.
The method of protein affinity modification by DNA analogues was used to study the characteristic features of restriction endonuclease MvaI interaction with DNA. Oligonucleotide duplexes containing a monosubstituted pyrophosphate internucleotide bond were used for cross-linking to the enzyme. The conditions of the reaction of MvaI endonuclease with these reagents were investigated. On the basis of data obtained, the model of successive inclusion of two Mg2+ ions into MvaI endonuclease-substrate complex was proposed and confirmed by the kinetic scheme of the process.
Sheflyan
G Y
GY
A.N. Belozersky Institute of Physical and Chemical Biology, Department of Chemistry, M.V. Lomonosov Moscow State University, 119899 Moscow, Russia.
Kubareva
E A
EA
Gromova
E S
ES
Shabarova
Z A
ZA
D000465:Algorithms; D003432:Cross-Linking Reagents; D004247:DNA / Q000737:chemistry*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry*; D007700:Kinetics; D008274:Magnesium / Q000737:chemistry*; D009841:Oligonucleotides / Q000737:chemistry; D011487:Protein Conformation
D003432:Cross-Linking Reagents; D009841:Oligonucleotides; D004247:DNA; C046545:CCWGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific; D008274:Magnesium