Matches in SemOpenAlex for { <https://semopenalex.org/work/W2014373216> ?p ?o ?g. }
Showing items 1 to 77 of
77
with 100 items per page.
- W2014373216 endingPage "67" @default.
- W2014373216 startingPage "61" @default.
- W2014373216 abstract "Lys146 of rabbit aldolase A [D-fructose-1,6-bis(phosphate): D-glyceraldehyde-3-phosphate lyase, EC 4.1.2.13] was changed to arginine by site-directed mutagenesis. The kcat of the resulting mutant protein, K146R, was 500 times slower than wild-type in steady-state kinetic assays for both cleavage and condensation of fructose-1,6-bis(phosphate), while the Km for this substrate was unchanged. Analysis of the rate of formation of catalytic intermediates showed K146R was significantly different from the wild-type enzyme and other enzymes mutated at this site. Single-turnover experiments using acid precipitation to trap the Schiff base intermediate on the wild-type enzyme failed to show a build-up of this intermediate on K146R. However, K146R retained the ability to form the Schiff base intermediate as shown by the significant amounts of Schiff base intermediate trapped with NaBH4. In the single-turnover experiments it appeared that the Schiff base intermediate was converted to products more rapidly than it was produced. This suggested a maximal rate of Schiff base formation of 0.022 s−1, which was close to the value of kcat for this enzyme. This observation is strikingly different from the wild-type enzyme in which Schiff base formation is >100 times faster than kcat. For K146R it appears that steps up to and including Schiff base formation are rate limiting for the catalytic reaction. The carbanion intermediate derived from either substrate or product, and the equilibrium concentrations of covalent enzyme-substrate intermediates, were much lower on K146R than on the wild-type enzyme. The greater bulk of the guanidino moiety may destabilize the covalent enzyme-substrate intermediates, thereby slowing the rate of Schiff base formation such that it becomes rate limiting. The K146R mutant enzyme is significantly more active than other enzymes mutated at this site, perhaps because it maintains a positively charged group at an essential position in the active site or perhaps the Arg functionally substitutes as a general acid/base catalyst in both Schiff base formation and in subsequent abstraction of the C4-hydroxyl proton." @default.
- W2014373216 created "2016-06-24" @default.
- W2014373216 creator A5027073083 @default.
- W2014373216 creator A5045614025 @default.
- W2014373216 creator A5059209346 @default.
- W2014373216 date "1996-01-01" @default.
- W2014373216 modified "2023-10-16" @default.
- W2014373216 title "A lysine to arginine substitution at position 146 of rabbit aldolase A changes the rate-determining step to Schiff base formation" @default.
- W2014373216 cites W1889077695 @default.
- W2014373216 cites W2144634347 @default.
- W2014373216 cites W404568673 @default.
- W2014373216 doi "https://doi.org/10.1093/protein/9.1.61" @default.
- W2014373216 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/9053904" @default.
- W2014373216 hasPublicationYear "1996" @default.
- W2014373216 type Work @default.
- W2014373216 sameAs 2014373216 @default.
- W2014373216 citedByCount "26" @default.
- W2014373216 countsByYear W20143732162013 @default.
- W2014373216 countsByYear W20143732162016 @default.
- W2014373216 countsByYear W20143732162021 @default.
- W2014373216 countsByYear W20143732162022 @default.
- W2014373216 countsByYear W20143732162023 @default.
- W2014373216 crossrefType "journal-article" @default.
- W2014373216 hasAuthorship W2014373216A5027073083 @default.
- W2014373216 hasAuthorship W2014373216A5045614025 @default.
- W2014373216 hasAuthorship W2014373216A5059209346 @default.
- W2014373216 hasBestOaLocation W20143732161 @default.
- W2014373216 hasConcept C181199279 @default.
- W2014373216 hasConcept C185592680 @default.
- W2014373216 hasConcept C18903297 @default.
- W2014373216 hasConcept C2776016237 @default.
- W2014373216 hasConcept C2776428424 @default.
- W2014373216 hasConcept C2777289219 @default.
- W2014373216 hasConcept C28165981 @default.
- W2014373216 hasConcept C41183919 @default.
- W2014373216 hasConcept C515207424 @default.
- W2014373216 hasConcept C55493867 @default.
- W2014373216 hasConcept C56856141 @default.
- W2014373216 hasConcept C63338738 @default.
- W2014373216 hasConcept C71240020 @default.
- W2014373216 hasConcept C86803240 @default.
- W2014373216 hasConceptScore W2014373216C181199279 @default.
- W2014373216 hasConceptScore W2014373216C185592680 @default.
- W2014373216 hasConceptScore W2014373216C18903297 @default.
- W2014373216 hasConceptScore W2014373216C2776016237 @default.
- W2014373216 hasConceptScore W2014373216C2776428424 @default.
- W2014373216 hasConceptScore W2014373216C2777289219 @default.
- W2014373216 hasConceptScore W2014373216C28165981 @default.
- W2014373216 hasConceptScore W2014373216C41183919 @default.
- W2014373216 hasConceptScore W2014373216C515207424 @default.
- W2014373216 hasConceptScore W2014373216C55493867 @default.
- W2014373216 hasConceptScore W2014373216C56856141 @default.
- W2014373216 hasConceptScore W2014373216C63338738 @default.
- W2014373216 hasConceptScore W2014373216C71240020 @default.
- W2014373216 hasConceptScore W2014373216C86803240 @default.
- W2014373216 hasIssue "1" @default.
- W2014373216 hasLocation W20143732161 @default.
- W2014373216 hasLocation W20143732162 @default.
- W2014373216 hasOpenAccess W2014373216 @default.
- W2014373216 hasPrimaryLocation W20143732161 @default.
- W2014373216 hasRelatedWork W1987826897 @default.
- W2014373216 hasRelatedWork W2003690817 @default.
- W2014373216 hasRelatedWork W2014373216 @default.
- W2014373216 hasRelatedWork W2051390995 @default.
- W2014373216 hasRelatedWork W2237717580 @default.
- W2014373216 hasRelatedWork W2317084518 @default.
- W2014373216 hasRelatedWork W2492855708 @default.
- W2014373216 hasRelatedWork W2494851022 @default.
- W2014373216 hasRelatedWork W2963115717 @default.
- W2014373216 hasRelatedWork W3019502410 @default.
- W2014373216 hasVolume "9" @default.
- W2014373216 isParatext "false" @default.
- W2014373216 isRetracted "false" @default.
- W2014373216 magId "2014373216" @default.
- W2014373216 workType "article" @default.