3.4.21.27 C147S inactive 754516 3.4.21.27 C321S the mutant shows wild type activity 754516 3.4.21.27 C362S/C482S site-directed mutagenesis 683682 3.4.21.27 C362S/C482S site-directed mutagenesis, the mutant shows increased platelet binding compared to the wild-type enzyme 683160 3.4.21.27 C398Y exhibits a dominant negative effect when coexpressed with wild-type FXI and is associated with FXI deficiency that may be inherited in a dominant manner 707856 3.4.21.27 C482S the mutant shows 1.45fold enhanced activity as compared with the wild type enzyme 754516 3.4.21.27 C500S inactive 731076 3.4.21.27 E98A the mutant has normal Km and kcat values for L-pyroGlu-L-Pro-L-Arg-4-nitroanilide hydrolysis, the mutant has deficient kcat values for factor IX hydrolysis, and is inhibited by protease nexin 2 with normal value of Ki 717844 3.4.21.27 E98D the mutant has normal Km and decreased kcat values for L-pyroGlu-L-Pro-L-Arg-4-nitroanilide hydrolysis, the mutant has deficient kcat values for factor IX hydrolysis 717844 3.4.21.27 E98V the mutant has increased Km and decreased kcat values for L-pyroGlu-L-Pro-L-Arg-4-nitroanilide hydrolysis, the mutant has deficient kcat values for factor IX hydrolysis 717844 3.4.21.27 FXI/G326C mutant of FXI with Gly326 residue mutated to Cys326 667620 3.4.21.27 G104R this mutation in the PK A2 domain associated with CRM+ PK deficiency causes decreased kininogen binding 707856 3.4.21.27 G155E is a rare example of a mutation causing CRM+ FXI deficieny 707856 3.4.21.27 G193A site-directed mutagenesis, the mutant shows reduced catalytic activity and impaired binding of inhibitors 4-aminobenzamidine and diisopropylfluorophosphate, respectively, indicating distortion of, or altered accessibility to, the S1 and oxyanion-binding sites 683169 3.4.21.27 G193D site-directed mutagenesis, the mutant shows reduced catalytic activity and binding of inhibitors 4-aminobenzamidine and diisopropylfluorophosphate impaired 1.6-36fold, respectively, indicating distortion of, or altered accessibility to, the S1 and oxyanion-binding sites 683169 3.4.21.27 G193E site-directed mutagenesis, the mutant shows reduced catalytic activity and impaired binding of inhibitors 4-aminobenzamidine and diisopropylfluorophosphate, respectively, indicating distortion of, or altered accessibility to, the S1 and oxyanion-binding sites 683169 3.4.21.27 G193K site-directed mutagenesis, the mutant shows reduced catalytic activity and binding of inhibitors 4-aminobenzamidine and diisopropylfluorophosphate impaired 35-478fold, respectively, indicating distortion of, or altered accessibility to, the S1 and oxyanion-binding sites 683169 3.4.21.27 G193R site-directed mutagenesis, the mutant shows reduced catalytic activity and impaired binding of inhibitors 4-aminobenzamidine and diisopropylfluorophosphate, respectively, indicating distortion of, or altered accessibility to, the S1 and oxyanion-binding sites 683169 3.4.21.27 G193V site-directed mutagenesis, the mutant shows reduced catalytic activity and impaired binding of inhibitors 4-aminobenzamidine and diisopropylfluorophosphate, respectively, indicating distortion of, or altered accessibility to, the S1 and oxyanion-binding sites 683169 3.4.21.27 G400V exhibits a dominant negative effect when coexpressed with wild-type FXI and is associated with FXI deficiency that may be inherited in a dominant manner 707856 3.4.21.27 I151A the mutant has normal Km and impaired kcat values for L-pyroGlu-L-Pro-L-Arg-4-nitroanilide hydrolysis, the mutant has deficient kcat values for factor IX hydrolysis, and is inhibited by protease nexin 2 with normal value of Ki 717844 3.4.21.27 K145A site-directed mutagenesis of an autolysis loop residue, the mutant shows altered sensitivity to inhibition by serpins compared to the wild-type enzyme 683157 3.4.21.27 K148A site-directed mutagenesis of an autolysis loop residue, the mutant shows altered sensitivity to inhibition by serpins compared to the wild-type enzyme 683157 3.4.21.27 K149A site-directed mutagenesis of an autolysis loop residue, the mutant shows altered sensitivity to inhibition by serpins compared to the wild-type enzyme 683157 3.4.21.27 K170A replaced the basic residues of the fXIa 170 loop (Lys-170, Arg-171, Arg-173, Lys-175, and Lys-179, chymotrypsin numbering) with Ala, using an expression system that allows separation of the fXIa catalytic domain from noncatalytic domains 691011 3.4.21.27 K170A/R171A/R173A catalytic domain with residues 170, 171, and 173 changed to alanine is designated CD-KRR/A 691011 3.4.21.27 K175A replaced the basic residues of the fXIa 170 loop (Lys-170, Arg-171, Arg-173, Lys-175, and Lys-179, chymotrypsin numbering) with Ala, using an expression system that allows separation of the fXIa catalytic domain from noncatalytic domains 691011 3.4.21.27 K179A replaced the basic residues of the fXIa 170 loop (Lys-170, Arg-171, Arg-173, Lys-175, and Lys-179, chymotrypsin numbering) with Ala, using an expression system that allows separation of the fXIa catalytic domain from noncatalytic domains 691011 3.4.21.27 K192A the mutant has normal Km value for L-pyroGlu-L-Pro-L-Arg-4-nitroanilide hydrolysis, the mutant has deficient kcat values for factor IX hydrolysis, and is inhibited by protease nexin 2 with increased value of Ki 717844 3.4.21.27 K192E the mutant has increased Km and decreased kcat values for L-pyroGlu-L-Pro-L-Arg-4-nitroanilide hydrolysis, the mutant has deficient kcat values for factor IX hydrolysis and is not inhibited by protease nexin 2 717844 3.4.21.27 K192Q the mutant has increased Km and decreased kcat values for L-pyroGlu-L-Pro-L-Arg-4-nitroanilide hydrolysis, the mutant has deficient kcat values for factor IX hydrolysis 717844 3.4.21.27 K192R the mutant has decreased Km and kcat values for L-pyroGlu-L-Pro-L-Arg-4-nitroanilide hydrolysis, the mutant has deficient kcat values for factor IX hydrolysis and is not inhibited by protease nexin 2 717844 3.4.21.27 M102T site-directed mutagenesis,missense mutation identified from large scale screening, the mutant enzyme is not secreted from the transfected cell resulting in a cross-reactive material negative phenotype 684078 3.4.21.27 M18I site-directed mutagenesis, missense mutation identified from large scale screening, the mutant enzyme is not secreted from the transfected cell resulting in a cross-reactive material negative phenotype 684078 3.4.21.27 additional information chimeric recombinant enzyme mutants rFXI-PKA3 and rFXIa-PKA3, with fusion of FXI to the Apple 3 domain of prekallikrein, show unaffected platelet binding compared to the wild-type enzyme, overview 683160 3.4.21.27 additional information construction of FXI with a single catalytic active site, construction of recombinant FXI with the A4 domain replaced by the A4 domain from plasma prekallikrein, i.e. FXI/PKA4, overview 683682 3.4.21.27 additional information factor XI deficiency caused by compound heterozygous F11 gene mutation 692544 3.4.21.27 additional information FXI with substitutions for Cys321 still form stable dimers. The Cys321-Cys321 interchain bond forms poorly in FXI with a single alanine substitution of Leu284, Ile290, or Tyr329 707856 3.4.21.27 additional information FXIa/PKA4 is made by replacing the FXI A4 domain with the A4 domain from prekallikrein. A dimeric version of FXI/PKA4 (FXI/PKA4-Gly326) is prepared as a control. Activated FXIa/PKA4 and FXIa/PKA4-Gly326 activate factor IX with kinetic parameters similar to that of FXIa. FXI/PKA4 and FXIa/PKA4-Gly326 have coagulant activity similar to FXI. FXIa/PKA4, FXIa/PKA4-Gly326 and FXIa have similar affinities for activated platelets. The dimeric proteins FXI and FXI/PKA4-Gly326 promote coagulation similarly, however, monomeric FXIa/PKA4 has greatly reduced activity. The activated monomeric FXIa/PKA4 activates factor IX poorly in the presence of activated platelets 650724 3.4.21.27 additional information mutations in the fXIa 170 helix are introduced into a modified human fXI cDNA (fXI-Ser-362,482), which contains serine substitutions for Cys-362 and Cys-482 691011 3.4.21.27 additional information two different mutations, c.1546 G>A (Val498Met) and c.1560dupG (Tyr503ValfsX32) in the F11 gene 691470 3.4.21.27 P520L site-directed mutagenesis, similar to the wild-ype enzyme 684078 3.4.21.27 R144A site-directed mutagenesis of an autolysis loop residue, the mutant shows altered sensitivity to inhibition by serpins compared to the wild-type enzyme 683157 3.4.21.27 R144A/K145A/R147A/K148A/K149A site-directed mutagenesis of an autolysis loop residues, the mutant shows altered sensitivity to inhibition by serpins compared to the wild-type enzyme 683157 3.4.21.27 R144A/K145A/R147A/R149A contains Ala substitutions for Arg-144, Lys-145, Arg-147, and Arg-149 (residues 504, 505, 507, and 509, respectively, in fXI numbering) 691011 3.4.21.27 R147A site-directed mutagenesis of an autolysis loop residue, the mutant shows altered sensitivity to inhibition by serpins compared to the wild-type enzyme 683157 3.4.21.27 R171A replaced the basic residues of the fXIa 170 loop (Lys-170, Arg-171, Arg-173, Lys-175, and Lys-179, chymotrypsin numbering) with Ala, using an expression system that allows separation of the fXIa catalytic domain from noncatalytic domains 691011 3.4.21.27 R173A replaced the basic residues of the fXIa 170 loop (Lys-170, Arg-171, Arg-173, Lys-175, and Lys-179, chymotrypsin numbering) with Ala, using an expression system that allows separation of the fXIa catalytic domain from noncatalytic domains 691011 3.4.21.27 R3704A the mutant has normal Km and impaired kcat values for L-pyroGlu-L-Pro-L-Arg-4-nitroanilide hydrolysis, the mutant has deficient kcat values for factor IX hydrolysis, and is inhibited by protease nexin 2 with normal value of Ki 717844 3.4.21.27 R378C site-directed mutagenesis, missense mutation identified from large scale screening, the mutant enzyme is normally secreted from the transfected cell, but shows negligible factor IX activation activity 684078 3.4.21.27 R37Q fXIa-R37Q mutant 691011 3.4.21.27 S225F exhibits a dominant negative effect when coexpressed with wild-type FXI and is associated with FXI deficiency that may be inherited in a dominant manner 707856 3.4.21.27 S248A binds platelets with reduced affinity compared with wild-type 707856 3.4.21.27 S248N binds platelets with 5fold reduced affinity compared with wild-type, the A3 domain is probably not affected significantly, as FXI-Asn248 is secreted, activated by activated factor XII, and activates FIX similar to wild-type activated factor IX. Is associated with bleeding and defective FXI binding to platelets but does not affect the activated partial thromboplastin time assay, which does not contain platelets 707856 3.4.21.27 S248Q binds platelets with reduced affinity compared with wild-type 707856 3.4.21.27 S434A/K437A/T475A/C482S to improve crystallizability a quadrupole mutant is generated 693547 3.4.21.27 S434A/T475A/C482S/K437A site-directed mutagenesis, crystal structure determination with bound benzylamidine 683756 3.4.21.27 T575M site-directed mutagenesis, missense mutation identified from large scale screening, the mutant enzyme is normally secreted from the transfected cell, but shows negligible factor IX activation activity 684078 3.4.21.27 W569S exhibits a dominant negative effect when coexpressed with wild-type FXI and is associated with FXI deficiency that may be inherited in a dominant manner 707856 3.4.21.27 Y133S site-directed mutagenesis, missense mutation identified from large scale screening, the mutant enzyme is not secreted from the transfected cell resulting in a cross-reactive material negative phenotype 684078 3.4.21.27 Y143A the mutant has normal Km and impaired kcat values for L-pyroGlu-L-Pro-L-Arg-4-nitroanilide hydrolysis, the mutant has deficient kcat values for factor IX hydrolysis, and is inhibited by protease nexin 2 with normal value of Ki 717844 3.4.21.27 Y5901A the mutant has normal Km and impaired kcat values for L-pyroGlu-L-Pro-L-Arg-4-nitroanilide hydrolysis, the mutant has deficient kcat values for factor IX hydrolysis, and is inhibited by protease nexin 2 with increased value of Ki 717844 3.4.21.27 Y5901V the mutant has normal Km and decreased kcat values for L-pyroGlu-L-Pro-L-Arg-4-nitroanilide hydrolysis, the mutant has deficient kcat values for factor IX hydrolysis and is not inhibited by protease nexin 2 717844