7.4.2.5 A123V partial loss of uptake 719940 7.4.2.5 A264P partial loss of uptake 719940 7.4.2.5 A285V complete loss of uptake 719940 7.4.2.5 A303G partial loss of uptake 719940 7.4.2.5 A68P complete loss of uptake 719940 7.4.2.5 coreTAP1 mutant, delta2-122 686718 7.4.2.5 coreTAP2 mutant, delta2-122 686718 7.4.2.5 Cys-less mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 D209A a dominant-negative mutant, binds ATP but is unable to hydrolyze it and is inactive in proOmpA translocation. Mutant generates a translocation intermediate of 18 kDa. Further addition of wild-type SecA causes its translocation into either mature OmpA or another intermediate of 28 kDa that can be translocated into mature by a proton motive force. The addition of excess D209N SecA during translocation causes a topology inversion of SecG 720044 7.4.2.5 D209N mutant of SecA capable of binding, but not hydrolysing ATP 688391 7.4.2.5 D630M ATPase activity is 0.6% of wild-type activity -, 667495 7.4.2.5 D668E mutation in the nucleotide-binding domain of TAP1 687520 7.4.2.5 D668E/Q701H TAP1 mutant 687533 7.4.2.5 D668E/Q710H mutation in the nucleotide-binding domain of TAP1 687520 7.4.2.5 D668N TAP1 mutant 687533 7.4.2.5 D668N/Q701A TAP1 mutant 687533 7.4.2.5 DELTAN1-2TAP1 mutant, delta2-89 686718 7.4.2.5 DELTAN1-2TAP2 mutant, delta2-44 686718 7.4.2.5 DELTAN1-3TAP1 mutant, delta2-132 686718 7.4.2.5 DELTAN1TAP1 mutant, delta2-44 686718 7.4.2.5 DELTAN1TAP2 mutant, delta2-44 686718 7.4.2.5 E20D mutant is not affected by the bulk pH in the range tested, no dramatic change in IC50 value for peptides Gly-Lys, beta-Ala-Lys 720986 7.4.2.5 E20Q mutant is not affected by the bulk pH in the range tested, no dramatic change in IC50 value for peptides Gly-Lys, beta-Ala-Lys 720986 7.4.2.5 E210Q mutant of SecA capable of binding, but not hydrolysing ATP 688391 7.4.2.5 E388D increase in IC50 value for peptides Gly-Lys, beta-Ala-Lys, increase in pH-optimum 720986 7.4.2.5 E388Q increase in IC50 value for peptides Gly-Lys, beta-Ala-Lys, increase in pH-optimum 720986 7.4.2.5 E400C mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 E400C/R642C mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 E400R mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 E400R/A628T mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 E400R/E619K mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 E400R/H620P mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 E400R/I627T mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 E400R/L610P mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 E400R/M607T mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 E400R/N629D mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 E485R/E488R/R367E mutation leads to a closed conformation of the clamp, the C-loop remains inside the clamp -, 734488 7.4.2.5 E56G complete loss of uptake 719940 7.4.2.5 E599Q 3000fold reduction of ATPase activity compared to wild-type 656145 7.4.2.5 E632D TAP2 mutant 687533 7.4.2.5 E632D/H661Q TAP2 mutant 687533 7.4.2.5 E632Q TAP2 mutant 687533 7.4.2.5 E632Q/H661A TAP2 mutant 687533 7.4.2.5 E632Q/H662A TAP2 mutant 687533 7.4.2.5 E635C mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 F197I change in selectivity 719940 7.4.2.5 F289L complete loss of uptake 719940 7.4.2.5 F289S complete loss of uptake 719940 7.4.2.5 F301I complete loss of uptake 719940 7.4.2.5 G101D partial loss of uptake 719940 7.4.2.5 G127D partial loss of uptake 719940 7.4.2.5 G55A mutation in the potential ATP-binding site in ExeA, decreased rate of aerolysin secretion 210378 7.4.2.5 G55D mutation in the potential ATP-binding site in ExeA, decreased rate of aerolysin secretion 210378 7.4.2.5 G55V mutation in the potential ATP-binding site in ExeA, decreased rate of aerolysin secretion 210378 7.4.2.5 G78C complete loss of uptake 719940 7.4.2.5 G86R partial loss of uptake 719940 7.4.2.5 H661Q TAP2 mutant 687533 7.4.2.5 H662A inactive 667693 7.4.2.5 I100V no membrane localization 719940 7.4.2.5 I122N partial loss of uptake 719940 7.4.2.5 I60N complete loss of uptake 719940 7.4.2.5 Ile3A mutation completely blocks dimerization of SecA in 300 mM KCl buffer 734081 7.4.2.5 K108R mutant, defective in ATP binding and protein translocation in vitro, as well as biologically inactive in vivo 687421 7.4.2.5 K115A mutant, substitution of the conserved lysine in the Walker A motif eliminates ATP binding and affects the biological activity -, 687421 7.4.2.5 K115R mutant, substitution of the conserved lysine in the Walker A motif eliminates ATP binding and affects the biological activity -, 687421 7.4.2.5 K274I change in selectivity 719940 7.4.2.5 K508M ATPase activity is 1.3% of wild-type activity -, 667495 7.4.2.5 K539M introduction of a Walker A mutation in the haf-4 construct. The mutation in ATP-binding site of HAF-4 inhibits the maturation of granules as dominant-negative effects. The inactive form of HAF-4 does not rescue the phenotype 700101 7.4.2.5 K545A/H699A TAPL double mutant 687663 7.4.2.5 L136R complete loss of uptake 719940 7.4.2.5 L137H no membrane localization 719940 7.4.2.5 L190V change in selectivity 719940 7.4.2.5 L2A/I3A mutation does not substantially affect SecA dimerization 734081 7.4.2.5 L324V change in selectivity 719940 7.4.2.5 L98R complete loss of uptake 719940 7.4.2.5 Leu2A mutation completely blocks dimerization of SecA in 300 mM KCl buffer 734081 7.4.2.5 Leu5A mutation completely blocks dimerization of SecA in 300 mM KCl buffer 734081 7.4.2.5 Leu6A mutation completely blocks dimerization of SecA in 300 mM KCl buffer 734081 7.4.2.5 M154K change in selectivity 719940 7.4.2.5 M295K complete loss of uptake 719940 7.4.2.5 additional information a collection of 63 monocysteine mutants for the 901-aminoacid-residue SecA protein is generated for topological analysis of the protein 687665 7.4.2.5 additional information analysis of the function of SecA via mutants and SecA-SecA2-chimeras 698615 7.4.2.5 additional information analysis of the function of SecA1 and SecA2 via several mutants 699002 7.4.2.5 additional information analysis of the function of SecA2 via mutants and SecA-SecA2-chimeras 698615 7.4.2.5 additional information analysis of the function of the two-helix finger of the SecA ATPase via point mutations 700390 7.4.2.5 additional information construction of SecA N-terminal deletion mutants. The first small helix, the linker and part of the second helix (residues 2-22) are dispensable for SecA activity in complementing the growth of a SecA temperature-sensitive mutant. Deletions of N-terminal aminoacyl residues 23-25 result in severe progressive retardation of growth. A decrease of SecA activity caused by N-terminal deletions correlates to the loss of SecA membrane binding, formation of lipid-specific domains and channel activity. The N-terminal aminoacyl residues 23-25 play a critical role for SecA binding to membranes and the N-terminal limit of SecA for activity is at the 25th amino acid 733274 7.4.2.5 additional information deletion of SecA1 failed, unless a copy of the gene is provided in trans on a plasmid, thus SecA1 belongs to the housekeeping genes 695920 7.4.2.5 additional information deletion of SecA2 failed, unless a copy of the gene is provided in trans on a plasmid, thus SecA2 belongs to the housekeeping genes 695920 7.4.2.5 additional information isoform PTR6 is able to complement transport defects in a yeast ptr2 mutant strain 720761 7.4.2.5 additional information knockdown of the ABC transporter enzyme in WM793B cells by shRNA leads to increases the sensitivity of cells to doxorubicin and reduces the cell survival, overview 700137 7.4.2.5 additional information large in-frame deletion of SpaT, not translocation is interrupted, but proteolytic cleavage of the leader segment while the precurser resides in the cell wall -, 210373 7.4.2.5 additional information mutant pairs of TAP1/TAP2 with mutation in ABC signature motif, C-loop. Chimera with two canonical C-loops show highest transport activity, chimera with two degenerate C-loops have lowest transport rates. All single mutants and chimera show similar activities in peptide or ATP binding 656253 7.4.2.5 additional information screening of a library of mutants to identify amino acids critical for peptide transport and identification of 35 single point mutations that result in a full or partial loss of transport activity 719940 7.4.2.5 additional information SecY-SecA interactions are studied by an in vivo site-directed cross-linking technique 689740 7.4.2.5 additional information the loss-of-function mutants for haf-4 and haf-9, constructed by RNAi mechanism, exhibit granular defects in late larval and young adult intestinal cells, associated with decreased brood size, prolonged defecation cycle, and slow growth. The intestinal granular phenotype is rescued by the overexpression of the GFP-tagged wild-type protein, but not by the ATP-unbound form of HAF-4, overview 700101 7.4.2.5 N196K partial loss of uptake 719940 7.4.2.5 N300I complete loss of uptake 719940 7.4.2.5 N300Y partial loss of uptake 719940 7.4.2.5 N306I partial loss of uptake 719940 7.4.2.5 N95 truncated version of Escherichia coli SecA, the last 70 residues are lacking 686725 7.4.2.5 N95CC two cysteines are introduced into a truncated version of Escherichia coli SecA, at position 636 and 801, the last 70 residues are lacking, mutant is dimeric and fully functional 686725 7.4.2.5 P326Q partial loss of uptake 719940 7.4.2.5 P624C insoluble mutant protein 667495 7.4.2.5 P624L insoluble mutant protein -, 667495 7.4.2.5 P624R insoluble mutant protein 667495 7.4.2.5 P624S insoluble mutant protein -, 667495 7.4.2.5 PaSecAL43P mutant 689856 7.4.2.5 PaSecAN236 N-terminal 236 amino acids of PaSecA 689856 7.4.2.5 Phe10A mutation completely blocks dimerization of SecA in 300 mM KCl buffer 734081 7.4.2.5 Q320L complete loss of uptake 719940 7.4.2.5 Q710H mutation in the nucleotide-binding domain of TAP1 687520 7.4.2.5 R305C partial loss of uptake 719940 7.4.2.5 R367E mutation of a conserved residue, results in significant lateral opening of the clamp, which leads to increased dissociation of the substrate -, 734488 7.4.2.5 R400R/M607T mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 R642C mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 R642E mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 R642E/A628T mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 R642E/E619K mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 R642E/H620P mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 R642E/I627T mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 R642E/L610P mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 R642E/M607T mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 R642E/N629D mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 R656C mutant for analyzing the interaction and regulatory domains of SecA 687527 7.4.2.5 S59P complete loss of uptake 719940 7.4.2.5 SecA2DELTA1-990 mutant, cloned into different plamids, construction of different strains 686782 7.4.2.5 SecA2DELTA1732-2397 7 mutant, cloned into different plamids, construction of different strains 686782 7.4.2.5 SecA2DELTA1768-2028 mutant, cloned into different plamids, construction of different strains 686782 7.4.2.5 SecA2DELTA991-1749 mutant, cloned into different plamids, construction of different strains 686782 7.4.2.5 SecAA tandem SecA 687406 7.4.2.5 SecADELTA11/N95 monomeric SecA derivative mutant, which lacks extreme terminal residues and shows in vitro and in vivo activities 687406 7.4.2.5 T297A complete loss of uptake 719940 7.4.2.5 TAP2DELTATM1 mutant, amino acids 2-42 are removed 687508 7.4.2.5 TAP2DELTATM2 mutant, amino acids 2-88 are removed 687508 7.4.2.5 TAP2DELTATM3 mutant, amino acids 2-128 are removed 687508 7.4.2.5 TAP2DELTATM4 mutant, amino acids 2-185 are removed 687508 7.4.2.5 TAP2DELTATM5 mutant, amino acids 2-241 are removed 687508 7.4.2.5 V252E change in selectivity 719940 7.4.2.5 V548A insoluble mutant protein -, 667495 7.4.2.5 V9A/F10A mutation enhances dissociation by 8fold with respect to that of wild-type SecA 734081 7.4.2.5 Val9A mutation completely blocks dimerization of SecA in 300 mM KCl buffer 734081 7.4.2.5 Y477W KM-value for ATP is 1.75fold higher than wild-type value. kcat for ATP is 4.3fold higher than wild-type value 667693