FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2060810897> ?p ?o ?g. }

• W2060810897 endingPage "22228" @default.
• W2060810897 startingPage "22223" @default.
• W2060810897 abstract "The serine residue required for catalysis of γ-glutamyl transpeptidase was identified by site-specific mutagenesis of the conserved serine residues on the basis of sequence alignment of the light subunit of human, rat, pig and two bacterial enzymes. Recombinant human γ-glutamyl transpeptidases with replacements of these serine residues by Ala were expressed using a baculovirus-insect cell system. Substitutions of Ala at Ser-385, −413 or −425 yielded almost fully active enzymes. However, substitutions of Ala at Ser-451 or −452 yielded enzymes that were only about 1% as active as the wild-type enzyme. Further, their double mutant is only 0.002% as active as the wild type. Kinetic analysis of transpeptidation using glycylglycine as acceptor indicates that the Vmax values of Ser-451 and −452 mutants are substantially decreased (to about 3% of the wild type); however, their Km values for L-γ-glutamyl-p-nitroanilide as donor were only increased about 5 fold compared to that of the wild type. The double mutation of Ser-451 and −452 further decreased the Vmax value to only about 0.005% of the wild type, while this mutation produced only a minor effect (2-fold increase) on the Km value for the donor. The kinetic values for the hydrolysis reaction of L-γ-glutamyl-p-nitroanilide in the mutants followed similar trends to those for transpeptidation. The rates of inactivation of Ser-451, −452 and their double mutant enzymes by acivicin, a potent inhibitor, were less than 1% that of the wild-type enzyme. The Ki value of the double mutant for L-serine as a competitive inhibitor of the γ-glutamyl group is only 9 fold increased over that of the wild type, whereas the Ki for the serine-borate complex, which acts as an inhibitory transition-state analog, was more than 1,000 times higher than for the wild-type enzyme. These results suggest that both Ser-451 and −452 are located at the position able to interact with the γ-glutamyl group and participate in catalysis, probably as nucleophiles or through stabilization of the transition state. The serine residue required for catalysis of γ-glutamyl transpeptidase was identified by site-specific mutagenesis of the conserved serine residues on the basis of sequence alignment of the light subunit of human, rat, pig and two bacterial enzymes. Recombinant human γ-glutamyl transpeptidases with replacements of these serine residues by Ala were expressed using a baculovirus-insect cell system. Substitutions of Ala at Ser-385, −413 or −425 yielded almost fully active enzymes. However, substitutions of Ala at Ser-451 or −452 yielded enzymes that were only about 1% as active as the wild-type enzyme. Further, their double mutant is only 0.002% as active as the wild type. Kinetic analysis of transpeptidation using glycylglycine as acceptor indicates that the Vmax values of Ser-451 and −452 mutants are substantially decreased (to about 3% of the wild type); however, their Km values for L-γ-glutamyl-p-nitroanilide as donor were only increased about 5 fold compared to that of the wild type. The double mutation of Ser-451 and −452 further decreased the Vmax value to only about 0.005% of the wild type, while this mutation produced only a minor effect (2-fold increase) on the Km value for the donor. The kinetic values for the hydrolysis reaction of L-γ-glutamyl-p-nitroanilide in the mutants followed similar trends to those for transpeptidation. The rates of inactivation of Ser-451, −452 and their double mutant enzymes by acivicin, a potent inhibitor, were less than 1% that of the wild-type enzyme. The Ki value of the double mutant for L-serine as a competitive inhibitor of the γ-glutamyl group is only 9 fold increased over that of the wild type, whereas the Ki for the serine-borate complex, which acts as an inhibitory transition-state analog, was more than 1,000 times higher than for the wild-type enzyme. These results suggest that both Ser-451 and −452 are located at the position able to interact with the γ-glutamyl group and participate in catalysis, probably as nucleophiles or through stabilization of the transition state." @default.
• W2060810897 created "2016-06-24" @default.
• W2060810897 creator A5000178422 @default.
• W2060810897 creator A5015017352 @default.
• W2060810897 creator A5044178319 @default.
• W2060810897 creator A5066654204 @default.
• W2060810897 creator A5090272663 @default.
• W2060810897 date "1995-09-01" @default.
• W2060810897 modified "2023-09-28" @default.
• W2060810897 title "Involvement of Ser-451 and Ser-452 in the Catalysis of Human γ-Glutamyl Transpeptidase" @default.
• W2060810897 cites W106439688 @default.
• W2060810897 cites W1223981148 @default.
• W2060810897 cites W1494935948 @default.
• W2060810897 cites W1501941897 @default.
• W2060810897 cites W1547761546 @default.
• W2060810897 cites W1604298442 @default.
• W2060810897 cites W1633567371 @default.
• W2060810897 cites W1673986517 @default.
• W2060810897 cites W1900191660 @default.
• W2060810897 cites W1971290197 @default.
• W2060810897 cites W1975488965 @default.
• W2060810897 cites W1975763435 @default.
• W2060810897 cites W1976479314 @default.
• W2060810897 cites W1995518418 @default.
• W2060810897 cites W1996448872 @default.
• W2060810897 cites W1998193327 @default.
• W2060810897 cites W2007224073 @default.
• W2060810897 cites W2007767207 @default.
• W2060810897 cites W2008405285 @default.
• W2060810897 cites W2008416160 @default.
• W2060810897 cites W2018093699 @default.
• W2060810897 cites W2019381912 @default.
• W2060810897 cites W2022961230 @default.
• W2060810897 cites W2023748923 @default.
• W2060810897 cites W2025957822 @default.
• W2060810897 cites W2036589422 @default.
• W2060810897 cites W2036931150 @default.
• W2060810897 cites W2042804712 @default.
• W2060810897 cites W2043663289 @default.
• W2060810897 cites W2061561132 @default.
• W2060810897 cites W2073125764 @default.
• W2060810897 cites W2075004701 @default.
• W2060810897 cites W2091748192 @default.
• W2060810897 cites W2100837269 @default.
• W2060810897 cites W2121437105 @default.
• W2060810897 cites W2138270253 @default.
• W2060810897 cites W4232534952 @default.
• W2060810897 cites W969775097 @default.
• W2060810897 doi "https://doi.org/10.1074/jbc.270.38.22223" @default.
• W2060810897 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/7673200" @default.
• W2060810897 hasPublicationYear "1995" @default.
• W2060810897 type Work @default.
• W2060810897 sameAs 2060810897 @default.
• W2060810897 citedByCount "64" @default.
• W2060810897 countsByYear W20608108972012 @default.
• W2060810897 countsByYear W20608108972013 @default.
• W2060810897 countsByYear W20608108972014 @default.
• W2060810897 countsByYear W20608108972015 @default.
• W2060810897 countsByYear W20608108972018 @default.
• W2060810897 countsByYear W20608108972019 @default.
• W2060810897 countsByYear W20608108972020 @default.
• W2060810897 countsByYear W20608108972021 @default.
• W2060810897 countsByYear W20608108972023 @default.
• W2060810897 crossrefType "journal-article" @default.
• W2060810897 hasAuthorship W2060810897A5000178422 @default.
• W2060810897 hasAuthorship W2060810897A5015017352 @default.
• W2060810897 hasAuthorship W2060810897A5044178319 @default.
• W2060810897 hasAuthorship W2060810897A5066654204 @default.
• W2060810897 hasAuthorship W2060810897A5090272663 @default.
• W2060810897 hasBestOaLocation W20608108971 @default.
• W2060810897 hasConcept C103408237 @default.
• W2060810897 hasConcept C104317684 @default.
• W2060810897 hasConcept C143065580 @default.
• W2060810897 hasConcept C153911025 @default.
• W2060810897 hasConcept C16318435 @default.
• W2060810897 hasConcept C181199279 @default.
• W2060810897 hasConcept C185592680 @default.
• W2060810897 hasConcept C207583985 @default.
• W2060810897 hasConcept C2776414213 @default.
• W2060810897 hasConcept C2777756961 @default.
• W2060810897 hasConcept C2780139244 @default.
• W2060810897 hasConcept C40767141 @default.
• W2060810897 hasConcept C41183919 @default.
• W2060810897 hasConcept C515207424 @default.
• W2060810897 hasConcept C55493867 @default.
• W2060810897 hasConcept C56856141 @default.
• W2060810897 hasConcept C71240020 @default.
• W2060810897 hasConcept C86803240 @default.
• W2060810897 hasConceptScore W2060810897C103408237 @default.
• W2060810897 hasConceptScore W2060810897C104317684 @default.
• W2060810897 hasConceptScore W2060810897C143065580 @default.
• W2060810897 hasConceptScore W2060810897C153911025 @default.
• W2060810897 hasConceptScore W2060810897C16318435 @default.
• W2060810897 hasConceptScore W2060810897C181199279 @default.
• W2060810897 hasConceptScore W2060810897C185592680 @default.
• W2060810897 hasConceptScore W2060810897C207583985 @default.
• W2060810897 hasConceptScore W2060810897C2776414213 @default.
• W2060810897 hasConceptScore W2060810897C2777756961 @default.

• next