Matches in SemOpenAlex for { <https://semopenalex.org/work/W2023248734> ?p ?o ?g. }
- W2023248734 endingPage "326" @default.
- W2023248734 startingPage "320" @default.
- W2023248734 abstract "The fragile histidine triad (Fhit) protein is a homodimeric protein with diadenosine 5',5'-P(1),P(3)-triphosphate (Ap(3)A) asymmetrical hydrolase activity. We have cloned the human cDNA Fhit in the pPROEX-1 vector and expressed with high yield in Escherichia coli with the sequence Met-Gly-His(6)-Asp-Tyr-Asp-Ile-Pro-Thr-Thr followed by a rTEV protease cleavage site, denoted as H6TV, fused to the N-terminus of Fhit. Expression of H6TV-Fhit in BL21(DE3) cells for 3 h at 37 degrees C produced 30 mg of H6TV-Fhit from 1 L of cell culture ( approximately 4 g of cells). The H6TV-Fhit protein was purified to homogeneity in a single step, with a yield of 80%, using nickel-nitrilotriacetate resin and imidazole buffer as eluting agent. Incubation of H6TV-Fhit with rTEV protease at 4 degrees C for 24 h resulted in complete cleavage of the H6TV peptide. There were no unspecific cleavage products. The purified Fhit protein could be stored for 3 weeks at 4 degrees C without loss of activity. The pure protein was stable at -20 degrees C for at least 18 months when stored in buffer containing 25% glycerol. Purified Fhit was highly active, with a K(m) value for Ap(3)A of 0.9 microM and a k(cat)(monomer) value of 7.2 +/- 1.6 s(-1) (n = 5). The catalytic properties of unconjugated Fhit protein and the H6TV-Fhit fusion protein were essentially identical. This indicates that the 24-amino-acid peptide containing the six histidines fused to the N-terminus of Fhit does not interfere in forming the active homodimers or in the binding of Ap(3)A." @default.
- W2023248734 created "2016-06-24" @default.
- W2023248734 creator A5007057665 @default.
- W2023248734 creator A5024607799 @default.
- W2023248734 creator A5072250438 @default.
- W2023248734 creator A5082550013 @default.
- W2023248734 date "2000-04-01" @default.
- W2023248734 modified "2023-10-18" @default.
- W2023248734 title "Expression in Escherichia coli and Simple Purification of Human Fhit Protein" @default.
- W2023248734 cites W1140394572 @default.
- W2023248734 cites W1967064347 @default.
- W2023248734 cites W1969195197 @default.
- W2023248734 cites W1969418387 @default.
- W2023248734 cites W1987719931 @default.
- W2023248734 cites W1989951007 @default.
- W2023248734 cites W1999318754 @default.
- W2023248734 cites W2002035803 @default.
- W2023248734 cites W2005642753 @default.
- W2023248734 cites W2015965147 @default.
- W2023248734 cites W2016794832 @default.
- W2023248734 cites W2061445402 @default.
- W2023248734 cites W2069951219 @default.
- W2023248734 cites W2084899119 @default.
- W2023248734 cites W2095058079 @default.
- W2023248734 cites W2100837269 @default.
- W2023248734 cites W2101524123 @default.
- W2023248734 cites W2126904200 @default.
- W2023248734 cites W2129423962 @default.
- W2023248734 cites W2323087188 @default.
- W2023248734 cites W2344414296 @default.
- W2023248734 cites W4293247451 @default.
- W2023248734 doi "https://doi.org/10.1006/prep.1999.1199" @default.
- W2023248734 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/10733886" @default.
- W2023248734 hasPublicationYear "2000" @default.
- W2023248734 type Work @default.
- W2023248734 sameAs 2023248734 @default.
- W2023248734 citedByCount "15" @default.
- W2023248734 countsByYear W20232487342012 @default.
- W2023248734 countsByYear W20232487342015 @default.
- W2023248734 crossrefType "journal-article" @default.
- W2023248734 hasAuthorship W2023248734A5007057665 @default.
- W2023248734 hasAuthorship W2023248734A5024607799 @default.
- W2023248734 hasAuthorship W2023248734A5072250438 @default.
- W2023248734 hasAuthorship W2023248734A5082550013 @default.
- W2023248734 hasConcept C104317684 @default.
- W2023248734 hasConcept C123894998 @default.
- W2023248734 hasConcept C151730666 @default.
- W2023248734 hasConcept C153911025 @default.
- W2023248734 hasConcept C175156509 @default.
- W2023248734 hasConcept C181199279 @default.
- W2023248734 hasConcept C185592680 @default.
- W2023248734 hasConcept C187882448 @default.
- W2023248734 hasConcept C2776714187 @default.
- W2023248734 hasConcept C2778264664 @default.
- W2023248734 hasConcept C2778460671 @default.
- W2023248734 hasConcept C2780646079 @default.
- W2023248734 hasConcept C33161422 @default.
- W2023248734 hasConcept C40767141 @default.
- W2023248734 hasConcept C43369102 @default.
- W2023248734 hasConcept C547475151 @default.
- W2023248734 hasConcept C55493867 @default.
- W2023248734 hasConcept C555283112 @default.
- W2023248734 hasConcept C86803240 @default.
- W2023248734 hasConceptScore W2023248734C104317684 @default.
- W2023248734 hasConceptScore W2023248734C123894998 @default.
- W2023248734 hasConceptScore W2023248734C151730666 @default.
- W2023248734 hasConceptScore W2023248734C153911025 @default.
- W2023248734 hasConceptScore W2023248734C175156509 @default.
- W2023248734 hasConceptScore W2023248734C181199279 @default.
- W2023248734 hasConceptScore W2023248734C185592680 @default.
- W2023248734 hasConceptScore W2023248734C187882448 @default.
- W2023248734 hasConceptScore W2023248734C2776714187 @default.
- W2023248734 hasConceptScore W2023248734C2778264664 @default.
- W2023248734 hasConceptScore W2023248734C2778460671 @default.
- W2023248734 hasConceptScore W2023248734C2780646079 @default.
- W2023248734 hasConceptScore W2023248734C33161422 @default.
- W2023248734 hasConceptScore W2023248734C40767141 @default.
- W2023248734 hasConceptScore W2023248734C43369102 @default.
- W2023248734 hasConceptScore W2023248734C547475151 @default.
- W2023248734 hasConceptScore W2023248734C55493867 @default.
- W2023248734 hasConceptScore W2023248734C555283112 @default.
- W2023248734 hasConceptScore W2023248734C86803240 @default.
- W2023248734 hasIssue "3" @default.
- W2023248734 hasLocation W20232487341 @default.
- W2023248734 hasLocation W20232487342 @default.
- W2023248734 hasOpenAccess W2023248734 @default.
- W2023248734 hasPrimaryLocation W20232487341 @default.
- W2023248734 hasRelatedWork W2122212633 @default.
- W2023248734 hasRelatedWork W2348991363 @default.
- W2023248734 hasRelatedWork W2350146650 @default.
- W2023248734 hasRelatedWork W2359862199 @default.
- W2023248734 hasRelatedWork W2367085638 @default.
- W2023248734 hasRelatedWork W2380100330 @default.
- W2023248734 hasRelatedWork W2382285667 @default.
- W2023248734 hasRelatedWork W2389269354 @default.
- W2023248734 hasRelatedWork W2413798589 @default.
- W2023248734 hasRelatedWork W4309884888 @default.
- W2023248734 hasVolume "18" @default.