FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2149068078> ?p ?o ?g. }

• W2149068078 endingPage "251" @default.
• W2149068078 startingPage "242" @default.
• W2149068078 abstract "Pigeon liver malic enzyme was inactivated and cleaved at Asp141, Asp194, and Asp464 by the Cu2+-ascorbate system in acidic environment. Site-specific mutagenesis was performed at these putative metal-binding sites. Three point mutants, D141N, D194N, and D464N; three double mutants, D(141,194)N, D(194,464)N, and D(141,464)N; and a triple mutant, D(141,194,464)N; as well as the wild-type malic enzyme (WT) were successfully cloned and expressed in Escherichia coli cells. All recombinant enzymes, except the triple mutant, were purified to apparent homogeneity by successive Q-Sepharose and adenosine-2′,5′-bisphosphate-agarose columns. The mutants showed similar apparent Km,nadp values to that of the WT. The Km, Mal value was increased in the D141N and D194N mutants. The Km, Mnvalue, on the other hand, was increased only in the D141N mutant by 14-fold, corresponding to ∼ 1.6 kcal/mol for the Asp141-Mn2+ binding energy. Substrate inhibition by L-malate was only observed in WT, D464N, and D(141,464)N. Initial velocity experiments were performed to derive the various kinetic parameters. The possible interactions between Asp141, Asp 194, and Asp464 were analyzed by the double-mutation cycles and triple-mutation box. There are synergistic weakening interactions between Asp141 and Asp194 in the metal binding that impel the D(141,194)N double mutant to an overall specificity constant [kcat/(Kd, Mn Km, Mal Km, NADP)] at least four orders of magnitude smaller than the WT value. This difference corresponds to an increase of 6.38 kcal/mol energy barrier for the catalytic efficiency. Mutation at Asp464, on the other hand, has partial additivity on the mutations at Asp141 and Asp194. The overall specificity constants for the double mutants D(194,464)N and D(141,464)N or the triple mutant D(141,194,464)N were decreased by only 10- to 100-fold compared to the WT. These results strongly suggest the involvement of Asp141 in the Mn2+-L-malate binding for the pigeon liver malic enzyme. The Asp194 and Asp464, which may be oxidized by nonspecific binding of Cu2+, are involved in the Mn2+-L-malate binding or catalysis indirectly by modulating the binding affinity of Asp141 with the Mn2+." @default.
• W2149068078 created "2016-06-24" @default.
• W2149068078 creator A5004904484 @default.
• W2149068078 creator A5007785553 @default.
• W2149068078 creator A5012339565 @default.
• W2149068078 creator A5015682606 @default.
• W2149068078 creator A5040214720 @default.
• W2149068078 creator A5071867905 @default.
• W2149068078 date "2008-12-31" @default.
• W2149068078 modified "2023-10-17" @default.
• W2149068078 title "Characterization of the functional role of asp 141, asp 194, and asp464 residues in the Mn2+-l-malate binding of pigeon liver malic enzyme" @default.
• W2149068078 cites W1545833105 @default.
• W2149068078 cites W1571975921 @default.
• W2149068078 cites W1849348813 @default.
• W2149068078 cites W1968445740 @default.
• W2149068078 cites W1974988689 @default.
• W2149068078 cites W1977257727 @default.
• W2149068078 cites W1984966127 @default.
• W2149068078 cites W1986082735 @default.
• W2149068078 cites W2006809862 @default.
• W2149068078 cites W2008440654 @default.
• W2149068078 cites W2011127629 @default.
• W2149068078 cites W2011673896 @default.
• W2149068078 cites W2014819743 @default.
• W2149068078 cites W2015361598 @default.
• W2149068078 cites W2020083750 @default.
• W2149068078 cites W2030620605 @default.
• W2149068078 cites W2038127925 @default.
• W2149068078 cites W2038472420 @default.
• W2149068078 cites W2048591080 @default.
• W2149068078 cites W2053344700 @default.
• W2149068078 cites W2053390244 @default.
• W2149068078 cites W2056476754 @default.
• W2149068078 cites W2058771034 @default.
• W2149068078 cites W2064828746 @default.
• W2149068078 cites W2067526555 @default.
• W2149068078 cites W2079525106 @default.
• W2149068078 cites W2081745487 @default.
• W2149068078 cites W2083316920 @default.
• W2149068078 cites W2084462600 @default.
• W2149068078 cites W2084740998 @default.
• W2149068078 cites W2084884863 @default.
• W2149068078 cites W2085940055 @default.
• W2149068078 cites W2087281541 @default.
• W2149068078 cites W2088843615 @default.
• W2149068078 cites W2099121794 @default.
• W2149068078 cites W2107919301 @default.
• W2149068078 cites W2128635872 @default.
• W2149068078 cites W2138270253 @default.
• W2149068078 cites W2146293054 @default.
• W2149068078 cites W2164830519 @default.
• W2149068078 cites W2168730967 @default.
• W2149068078 cites W4247816627 @default.
• W2149068078 cites W4293247451 @default.
• W2149068078 cites W46798781 @default.
• W2149068078 cites W80140822 @default.
• W2149068078 doi "https://doi.org/10.1110/ps.9.2.242" @default.
• W2149068078 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/2144557" @default.
• W2149068078 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/10716176" @default.
• W2149068078 hasPublicationYear "2008" @default.
• W2149068078 type Work @default.
• W2149068078 sameAs 2149068078 @default.
• W2149068078 citedByCount "3" @default.
• W2149068078 crossrefType "journal-article" @default.
• W2149068078 hasAuthorship W2149068078A5004904484 @default.
• W2149068078 hasAuthorship W2149068078A5007785553 @default.
• W2149068078 hasAuthorship W2149068078A5012339565 @default.
• W2149068078 hasAuthorship W2149068078A5015682606 @default.
• W2149068078 hasAuthorship W2149068078A5040214720 @default.
• W2149068078 hasAuthorship W2149068078A5071867905 @default.
• W2149068078 hasBestOaLocation W21490680781 @default.
• W2149068078 hasConcept C103408237 @default.
• W2149068078 hasConcept C104317684 @default.
• W2149068078 hasConcept C107824862 @default.
• W2149068078 hasConcept C143065580 @default.
• W2149068078 hasConcept C153911025 @default.
• W2149068078 hasConcept C181199279 @default.
• W2149068078 hasConcept C185592680 @default.
• W2149068078 hasConcept C2776317432 @default.
• W2149068078 hasConcept C2778957662 @default.
• W2149068078 hasConcept C41183919 @default.
• W2149068078 hasConcept C547475151 @default.
• W2149068078 hasConcept C55493867 @default.
• W2149068078 hasConcept C56856141 @default.
• W2149068078 hasConcept C71240020 @default.
• W2149068078 hasConcept C86803240 @default.
• W2149068078 hasConceptScore W2149068078C103408237 @default.
• W2149068078 hasConceptScore W2149068078C104317684 @default.
• W2149068078 hasConceptScore W2149068078C107824862 @default.
• W2149068078 hasConceptScore W2149068078C143065580 @default.
• W2149068078 hasConceptScore W2149068078C153911025 @default.
• W2149068078 hasConceptScore W2149068078C181199279 @default.
• W2149068078 hasConceptScore W2149068078C185592680 @default.
• W2149068078 hasConceptScore W2149068078C2776317432 @default.
• W2149068078 hasConceptScore W2149068078C2778957662 @default.
• W2149068078 hasConceptScore W2149068078C41183919 @default.
• W2149068078 hasConceptScore W2149068078C547475151 @default.
• W2149068078 hasConceptScore W2149068078C55493867 @default.

• next