3.6.4.B7: RadA recombinase

RadA mutant C28Y retains the ATPase activity but is defective in DNA binding

K108R

the mutation at the Walker A sequence results in a dominant-negative RadA allele in Escherichia coli, it shows highly reduced DNA binding compared to wild-type

K258A

the mutation in the KNRFG motif negates RadA function and is partially dominant in vivo, it shows highly reduced DNA binding compared to wild-type

S372A

the RadA mutant retains ATPase and DNA-binding activities similar to wild-type

D246A

loss of binding a second Mg2+. Initial ATP turnover rate is reduced by about 20-fold

724274

D246N

initial ATP turnover rate is reduced by about 20-fold

724274

D302K

mutant protein shows comparable strand exchange efficiencies in the presence of either potassium or sodium

725699

E151D

mutant protein retains potassium preference in promoting strand exchange. Reduced ATPase activity and normal strand exchange activity

725699

E151K

mutant protein retains potassium preference in promoting strand exchange. Reduced ATPase activity and normal strand exchange activity

725699

E354A

site-directed mutagenesis, mutant M9, matuation of a residue involved in ATPase function, and coordination of the nucleophilic water molecule

E360A

site-directed mutagenesis, mutant M5, mutation of residues involved in exstein-intein interaction

E360A/R363A/E364A

site-directed mutagenesis, mutant M3, mutation of residues involved in exstein-intein interaction

E364A

site-directed mutagenesis, mutant M7, mutation of residues involved in exstein-intein interaction

E57A/K58A/R60A/E61A

site-directed mutagenesis, mutant M12, mutation of a residue that aisnot expected to be in proximity to intein catalytic residues

E77A/K79A/E80A

site-directed mutagenesis, mutant M11, mutation of a residue that aisnot expected to be in proximity to intein catalytic residues

I169M/Y201A/V202Y/E219S/D220A/K221M

site-directed mutagenesis, construction of mutant, termed HumRadA2, that resembles human RAD51. The mutant shows reduced thermal stability compared to wild-type, while the crystal structure of this mutant shows no structural changes in the ATP-binding site with respect to the wild-type. Fluorescence-based thermal shift and isothermal titration calorimetric analyses of nucleotide binding to recombinant mutant monomeric HumRadA2, overview

756730

Q465A

site-directed mutagenesis, mutant M10, matuation of a residue involved in ATPase function, and coordination of the nucleophilic water molecule

R358A

site-directed mutagenesis, mutant M4, mutation of residues involved in exstein-intein interaction

R358A/E360A/R363A/E364A

site-directed mutagenesis, mutant M1, mutation of residues involved in exstein-intein interaction

R358A/R361A

site-directed mutagenesis, mutant M2, mutation of residues involved in exstein-intein interaction

R361A

site-directed mutagenesis, mutant M6, mutation of residues involved in exstein-intein interaction

R503A

site-directed mutagenesis, mutant M8, mutation of residues involved in exstein-intein interaction

I169M/Y201A/V202Y/E219S/D220A/K221M

4 entries

E354A

site-directed mutagenesis, mutant M9, matuation of a residue involved in ATPase function, and coordination of the nucleophilic water molecule

E360A

site-directed mutagenesis, mutant M5, mutation of residues involved in exstein-intein interaction

I169M/Y201A/V202Y/E219S/D220A/K221M

R358A

site-directed mutagenesis, mutant M4, mutation of residues involved in exstein-intein interaction

R361A

site-directed mutagenesis, mutant M6, mutation of residues involved in exstein-intein interaction

R503A

site-directed mutagenesis, mutant M8, mutation of residues involved in exstein-intein interaction

K120A

reduced ATPase activity, mutant K120A is able to bind ssDNA with ATP, ADP, or ATPgammaS under saturating protein conditions, but failed to bind well at subsaturating concentrations with ATP or ATPgammaS

724820

K120R

reduced ATPase activity, mutant only binds ATP in the presence of ssDNA

724820

K27A

2 entries

K27R

mutant does not produced a D-loop product as compared to that of the wild type SsoRadA protein, exhibits weaker affinity to dsDNA as compared to wild-type protein

K60A

2 entries

K60R

mutant does not produced a D-loop product as compared to that of the wild type SsoRadA protein, binds dsDNA as well as wild-type protein

R217A

mutant does not produced a D-loop product as compared to that of the wild type SsoRadA protein, association and dissociation kinetics largely identical or similar to that of the wild-type protein, exhibits weaker affinity to dsDNA as compared to wild-type protein

R217K

mutant does not produced a D-loop product as compared to that of the wild type SsoRadA protein, mutant exhibits slower ssDNA association rate, surface plasmon resonance binding signals is similar to that of wild-type protein, exhibits weaker affinity to dsDNA as compared to wild-type protein

R223A

mutant does not produced a D-loop product as compared to that of the wild type SsoRadA protein, association and dissociation kinetics largely identical or similar to that of the wild-type protein, 90100% reduction of the surface plasmon resonance binding signal, mutants is defective in dsDNA binding

R223K

mutant does not produced a D-loop product as compared to that of the wild type SsoRadA protein, surface plasmon resonance binding signals is similar to that of wild-type protein, mutants is defective in dsDNA binding

R229A

mutant does not produced a D-loop product as compared to that of the wild type SsoRadA protein, association and dissociation kinetics largely identical or similar to that of the wild-type protein, 90100% reduction of the surface plasmon resonance binding signal, mutants is defective in dsDNA binding

R229K