7.1.1.1 A246C reverse activity stronger affected than cyclic activity 658003 7.1.1.1 A253C reverse activity stronger affected than cyclic activity 658003 7.1.1.1 A348C mutation introduced into a cysteine-free mutant enzyme, mutant shows markedly reduced activity 392663 7.1.1.1 A390C mutation introduced into a cysteine-free mutant enzyme 392663 7.1.1.1 A398C the mutant with wild type activity shows increased ratios between the rates of the forward and reverse reactions, thus approaching that of the wild type enzyme 733772 7.1.1.1 A432C mutation in domain III, reverse reaction in the presence of domain I from R. rubrum, 150% higher reaction rate than wild-type domain III/R. rubrum domain I mixture 392667 7.1.1.1 A432C the mutant shows increased ratios between the rates of the forward and reverse reactions, thus approaching that of the wild type enzyme 733772 7.1.1.1 C292T/C339T/C395S/C397T/C435S cysteine of the alpha subunits replaced, similar activity as wild-type 392662 7.1.1.1 C292T/C339T/C395S/C397T/C435S/C147S/C260S all 7 cysteines of the enzyme, 5 localized in the alpha subunit and 2 in the beta subunit, are replaced, the cysteine-free mutant shows about 5fold more activity in the reduction of acetylpyridine adenine dinucleotide by NADH than wild-type, the cyclic reduction of acetylpyridine adenine dinucleotide by NADH via NADPH is 2-2.5fold more activ 392662 7.1.1.1 D135N mutation has no effect in binding affinity of either NAD+ or NADH 674550 7.1.1.1 D135N the mutant shows reduced activity compared to the wild type enzyme 674550 7.1.1.1 D213K mutation in domain II 392671 7.1.1.1 D213R mutation in domain II 392671 7.1.1.1 D238C reverse activity stronger affected than cyclic activity 658003 7.1.1.1 D392C the mutant shows increased ratios between the rates of the forward and reverse reactions, thus approaching that of the wild type enzyme 733772 7.1.1.1 D401E mutation in beta subunit 392664 7.1.1.1 D401G mutation in beta subunit 392664 7.1.1.1 D401V mutation in beta subunit 392664 7.1.1.1 E124C domian dII, strongly reduced reverse activity, no effect on cyclic activity 658197 7.1.1.1 E155W dIII domain 657960 7.1.1.1 E155W dIII domain, catalytic properties are similar to the wild type dIII 658162 7.1.1.1 E155W mutant of the NADP(H)-binding component 392674 7.1.1.1 E155W mutation in domain III, similar catalytic activities as wild-type, used for tryptophan fluorescence measurements 392674 7.1.1.1 E155W the mutant shows reduced activity compared to the wild type enzyme 674550 7.1.1.1 E155W the mutant shows wild type activity 733378 7.1.1.1 E155W/G173C dIII domain, catalytic properties are similar to the wild type dIII, increased rate of reverse reaction 658162 7.1.1.1 E155W/R165A the mutations mutation do not significantly affect catalytic activity 733378 7.1.1.1 E413D mutation in beta subunit 392664 7.1.1.1 E413G mutation in beta subunit 392664 7.1.1.1 E413V mutation in beta subunit 392664 7.1.1.1 G132A in domain dII, no effect on wild type reverse activity 659274 7.1.1.1 G138A in domain dII, 57% of wild type reverse activity 659274 7.1.1.1 G226A in domain dII, 50% of wild type reverse activity 659274 7.1.1.1 G233A in domain dII, 49% of wild type reverse activity 659274 7.1.1.1 G245A in domain dII, no effect of wild type reverse activity 659274 7.1.1.1 G245C 24% of reverse activity 658003 7.1.1.1 G245L 52% of cyclic activity 658003 7.1.1.1 G245L the mutation leads to a general inhibition of all enzyme activities 658003 7.1.1.1 G249A in domain dII, 79% wild type reverse activity 659274 7.1.1.1 G249C 40% of reverse activity 658003 7.1.1.1 G249L 48% of cyclic activity 658003 7.1.1.1 G249L 70% of reverse activity 658003 7.1.1.1 G249L the mutation leads to a general inhibition of all enzyme activities 658003 7.1.1.1 G252A in domain dII, 2.6% of wild type reverse activity 659274 7.1.1.1 G252C in domain dII, 1.9% of wild type reverse activity 659274 7.1.1.1 G252C less than 5% of reverse activity 658003 7.1.1.1 G252L 13% of cyclic activity 658003 7.1.1.1 G252L the mutation leads to a general inhibition of all enzyme activities 658003 7.1.1.1 G252S in domain dII, 2.4% of wild type reverse activity 659274 7.1.1.1 G252T in domain dII, 2.3% of wild type reverse activity 659274 7.1.1.1 G252V in domain dII, 2.5% of wild type reverse activity 659274 7.1.1.1 G408C the mutant with wild type activity shows increased ratios between the rates of the forward and reverse reactions, thus approaching that of the wild type enzyme 733772 7.1.1.1 G430C mutation in domain III, reverse reaction in the presence of domain I from R. rubrum, 850% higher reaction rate than wild-type domain III/R. rubrum domain I mixture 392667 7.1.1.1 G430C the mutant shows increased ratios between the rates of the forward and reverse reactions, thus approaching that of the wild type enzyme 733772 7.1.1.1 G476C domian dII, little effect on activity 658197 7.1.1.1 G95A in domain dII, 56% of wild type reverse activity 659274 7.1.1.1 H91C the mutant of the beta subunit is unable to undergo the conformational change that occurs on binding of the substrates NADP+ or NADPH. The mutant retains 12% of the hydride transfer activity while proton translocation is reduced to 7% compared to the wild type enzyme 733317 7.1.1.1 H91D the mutant of the beta subunit retains 15% of the hydride transfer activity while proton translocation is reduced to 9% compared to the wild type enzyme 733317 7.1.1.1 H91E mutation in beta subunit 392664 7.1.1.1 H91K mutation in beta subunit 392664 7.1.1.1 H91K mutation in domain II, leads to occlusion of NADP(H) at the NADP(H)-binding site of domain III 392671 7.1.1.1 H91K the mutant of the beta subunit is present in the NADP(H)-induced conformation even in the absence of these substrates. The mutant retains 4% of the hydride transfer activity while proton translocation is reduced to 20% compared to the wild type enzyme 733317 7.1.1.1 H91N the mutant of the beta subunit retains 80% of the hydride transfer activity while proton translocation is reduced to 7% compared to the wild type enzyme 733317 7.1.1.1 H91R mutation in domain II, leads to occlusion of NADP(H) at the NADP(H)-binding site of domain III 392671 7.1.1.1 H91S the mutant of the beta subunit is unable to undergo the conformational change that occurs on binding of the substrates NADP+ or NADPH. The mutant retains 19% of the hydride transfer activity while proton translocation is reduced to 11% compared to the wild type enzyme 733317 7.1.1.1 H91T the mutant of the beta subunit is unable to undergo the conformational change that occurs on binding of the substrates NADP+ or NADPH. The mutant retains 11% of the hydride transfer activity while proton translocation is reduced to 8% compared to the wild type enzyme 733317 7.1.1.1 I258C reverse activity stronger affected than cyclic activity 658003 7.1.1.1 I406C the mutant with 450% of wild type activity shows increased ratios between the rates of the forward and reverse reactions, thus approaching that of the wild type enzyme 733772 7.1.1.1 K416G mutation in beta subunit 392664 7.1.1.1 K424C mutation introduced into a cysteine-free mutant enzyme, mutant shows markedly reduced activity 392663 7.1.1.1 K424G mutation in beta subunit 392664 7.1.1.1 K424R mutation in beta subunit 392664 7.1.1.1 K452D mutation in beta subunit 392664 7.1.1.1 K452G mutation in beta subunit 392664 7.1.1.1 L240C reverse activity stronger affected than cyclic activity 658003 7.1.1.1 L241C reverse activity stronger affected than cyclic activity 658003 7.1.1.1 L254C reverse activity stronger affected than cyclic activity 658003 7.1.1.1 L255C reverse activity stronger affected than cyclic activity 658003 7.1.1.1 M239F the Km for APAD+ during reverse transhydrogenation is 6fold greater compared to the wild type. Cyclic transhydrogenation (in membranes and the recombinant system) is substantially more inhibited (84%) than either forward or reverse transhydrogenation 733318 7.1.1.1 M259C 21% of reverse activity, 215% of cyclic activity 658003 7.1.1.1 M293I the Km for APAD+ during reverse transhydrogenation is 5fold greater compared to the wild type. Cyclic transhydrogenation (in membranes and the recombinant system) is substantially more inhibited (70%) than either forward or reverse transhydrogenation 733318 7.1.1.1 M409C the mutant with 75% of wild type activity shows increased ratios between the rates of the forward and reverse reactions, thus approaching that of the wild type enzyme 733772 7.1.1.1 additional information properties of a variety of mutant enzymes containing modified conserved and semiconserved basic and acidic residues in the beta subunit 392664 7.1.1.1 N222K mutation in domain II, leads to occlusion of NADP(H) at the NADP(H)-binding site of domain III 392671 7.1.1.1 N222R mutation in domain II, leads to occlusion of NADP(H) at the NADP(H)-binding site of domain III 392671 7.1.1.1 N238C reverse activity stronger affected than cyclic activity 658003 7.1.1.1 Q132N dI domain, no cyclic transhydrogenation activity in mixtures of domain dIII with the dI mutant, mutation has little effect on the NADH binding affinity 657960 7.1.1.1 Q132N the mutant shows reduced activity compared to the wild type enzyme 674550 7.1.1.1 R127A mutation strongly inhibits the rate of transhydrogenation and alters the nucleotide-binding properties of the dI protein. When dIR127A is reconstituted into the intact enzyme in membranes, transhydrogenation rates are negligible. dI is the NAD(H)-binding component of the transhydrogenase 674550 7.1.1.1 R127A the mutant shows reduced activity compared to the wild type enzyme 674550 7.1.1.1 R127M mutation strongly inhibits the rate of transhydrogenation and alters the nucleotide-binding properties of the dI protein. When dIR127M is reconstituted into the intact enzyme in membranes, transhydrogenation rates are negligible. dI is the NAD(H)-binding component of the transhydrogenase 674550 7.1.1.1 R127M the mutant shows reduced activity compared to the wild type enzyme 674550 7.1.1.1 R165A a higher concentration of the nucleotide is needed to achieve the half-maximal rate compared to with the wild type protein 733378 7.1.1.1 R425C mutation in domain III, reverse reaction in the presence of domain I from R. rubrum, 425% higher reaction rate than wild-type domain III/R. rubrum domain I mixture 392667 7.1.1.1 R425C mutation introduced into a cysteine-free mutant enzyme, mutant shows markedly reduced activity 392663 7.1.1.1 R425E mutation in beta subunit 392664 7.1.1.1 R425G mutation in beta subunit 392664 7.1.1.1 R425K mutation in beta subunit 392664 7.1.1.1 S105C domian dII, significantly reduced activity 658197 7.1.1.1 S135A mutation has no effect in binding affinity of either NAD+ or NADH 674550 7.1.1.1 S138A the mutant shows reduced activity compared to the wild type enzyme 674550 7.1.1.1 S183C domian dII, significantly reduced activity 658197 7.1.1.1 S237C domian dII, slightly reduced reverse activity, no efect on cyclic activity 658197 7.1.1.1 S250C strongly increased reverse and cyclic activity 658003 7.1.1.1 S250C the mutation leads to enhanced activities of all enzyme activities 658003 7.1.1.1 S251C strongly increased reverse and cyclic activity 658003 7.1.1.1 S251C the mutation leads to enhanced activities of all enzyme activities 658003 7.1.1.1 S256C the mutation leads to enhanced activities of all enzyme activities 658003 7.1.1.1 S2C domian dII, no effect on activity 658197 7.1.1.1 S404C the mutant with 75% of wild type activity shows increased ratios between the rates of the forward and reverse reactions, thus approaching that of the wild type enzyme 733772 7.1.1.1 T214A mutation in chain B, 3-5% residual activity -, 765844 7.1.1.1 T214A the mutant shows 5% or less of wild type activity -, 742240 7.1.1.1 T244C reverse activity stronger affected than cyclic activity 658003 7.1.1.1 T393C mutation in domain III, reverse reaction in the presence of domain I from R. rubrum, 175% higher reaction rate than wild-type domain III/R. rubrum domain I mixture 392667 7.1.1.1 T393C the mutant shows increased ratios between the rates of the forward and reverse reactions, thus approaching that of the wild type enzyme 733772 7.1.1.1 T54C domian dII, significantly reduced activity 658197 7.1.1.1 V243C reverse activity stronger affected than cyclic activity 658003 7.1.1.1 V248C reverse activity stronger affected than cyclic activity 658003 7.1.1.1 V411C the mutant with 125% of wild type activity shows increased ratios between the rates of the forward and reverse reactions, thus approaching that of the wild type enzyme 733772 7.1.1.1 W72F the mutant shows wild type activity 733378 7.1.1.1 Y146A mutation in component dI that binds NADH. dI.Y146A more readily dissociates into monomers than wild-type dI. dI.Y146A monomers bind NADH much more weakly than dimers. dI.Y146A reconstitutes activity to dI-depleted membranes in its dimeric form but not in its monomeric form 687656 7.1.1.1 Y146A the mutant binds NADH much more weakly than the wild type enzyme 687656 7.1.1.1 Y146F mutation in component dI that binds NADH. Wild-type dI and dI.Y146F reconstituted activity to dI-depleted membranes with similar characteristics 687656 7.1.1.1 Y146F the mutant shows wild type NADH binding ability 687656 7.1.1.1 Y171W mutation in domain III, similar catalytic activities as wild-type, used for tryptophan fluorescence measurements 392674 7.1.1.1 Y171W the mutant shows wild type activity 392674 7.1.1.1 Y257C reverse activity stronger affected than cyclic activity 658003 7.1.1.1 Y431C the mutant with 450% of wild type activity shows increased ratios between the rates of the forward and reverse reactions, thus approaching that of the wild type enzyme 733772