FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2008478712> ?p ?o ?g. }

• W2008478712 endingPage "20698" @default.
• W2008478712 startingPage "20690" @default.
• W2008478712 abstract "The τ subunit dimerizes DNA polymerase III via interaction with the α subunit, allowing DNA polymerase III holoenzyme to synthesize both leading and lagging strands simultaneously at the DNA replication fork. Here, we report a general method to map the limits of domains required for heterologous protein-protein interactions using surface plasmon resonance. The method employs fusion of a short biotinylation sequence at either the NH2 or COOH terminus of the protein to be immobilized on streptavidin-derivatized biosensor chips. Inclusion of a hexahistidine sequence permits rapid purification and separation of the fusion protein from the endogenous Escherichia coli biotin carboxyl carrier protein. Ten deletions of the α subunit were constructed and purified by Ni2+-nitrilotriacetic acid chromatography and, when required, monomeric avidin chromatography. Each α deletion protein was captured by streptavidin immobilized on a Pharmacia Biosensor BIAcore chip, and the τ binding activity of each α deletion was analyzed using surface plasmon resonance. The τ subunit bound very tightly to a full-length amino-terminal fusion of the biotinylation sequence with α (KD ∼ 70 pM). Four additional NH2-terminal α deletion proteins (60, 240, 360, and 542 residues deleted) retained strong binding activity to the τ subunit (KD = 0.19-0.39 nM), whereas deletion of 705 residues or more from the NH2 terminus of the α subunit abolished τ binding activity. Full-length α that contained a carboxyl-terminal fusion with the biotinylation sequence bound τ strongly (KD = 0.37 nM). However, deletion of 48 amino acids from the COOH terminus totally eliminated τ binding. These results indicate that the COOH-terminal half of the α subunit is involved in τ interaction. The τ subunit dimerizes DNA polymerase III via interaction with the α subunit, allowing DNA polymerase III holoenzyme to synthesize both leading and lagging strands simultaneously at the DNA replication fork. Here, we report a general method to map the limits of domains required for heterologous protein-protein interactions using surface plasmon resonance. The method employs fusion of a short biotinylation sequence at either the NH2 or COOH terminus of the protein to be immobilized on streptavidin-derivatized biosensor chips. Inclusion of a hexahistidine sequence permits rapid purification and separation of the fusion protein from the endogenous Escherichia coli biotin carboxyl carrier protein. Ten deletions of the α subunit were constructed and purified by Ni2+-nitrilotriacetic acid chromatography and, when required, monomeric avidin chromatography. Each α deletion protein was captured by streptavidin immobilized on a Pharmacia Biosensor BIAcore chip, and the τ binding activity of each α deletion was analyzed using surface plasmon resonance. The τ subunit bound very tightly to a full-length amino-terminal fusion of the biotinylation sequence with α (KD ∼ 70 pM). Four additional NH2-terminal α deletion proteins (60, 240, 360, and 542 residues deleted) retained strong binding activity to the τ subunit (KD = 0.19-0.39 nM), whereas deletion of 705 residues or more from the NH2 terminus of the α subunit abolished τ binding activity. Full-length α that contained a carboxyl-terminal fusion with the biotinylation sequence bound τ strongly (KD = 0.37 nM). However, deletion of 48 amino acids from the COOH terminus totally eliminated τ binding. These results indicate that the COOH-terminal half of the α subunit is involved in τ interaction." @default.
• W2008478712 created "2016-06-24" @default.
• W2008478712 creator A5005151768 @default.
• W2008478712 creator A5021046528 @default.
• W2008478712 date "1996-08-01" @default.
• W2008478712 modified "2023-10-16" @default.
• W2008478712 title "Biotin Tagging Deletion Analysis of Domain Limits Involved in Protein-Macromolecular Interactions" @default.
• W2008478712 cites W1484429384 @default.
• W2008478712 cites W1484791891 @default.
• W2008478712 cites W1508943098 @default.
• W2008478712 cites W1517901125 @default.
• W2008478712 cites W1531749456 @default.
• W2008478712 cites W1535233331 @default.
• W2008478712 cites W1542141332 @default.
• W2008478712 cites W1546339757 @default.
• W2008478712 cites W1548939450 @default.
• W2008478712 cites W1550211736 @default.
• W2008478712 cites W1551043626 @default.
• W2008478712 cites W1551254087 @default.
• W2008478712 cites W1560835792 @default.
• W2008478712 cites W1575133253 @default.
• W2008478712 cites W1575315612 @default.
• W2008478712 cites W1586315068 @default.
• W2008478712 cites W1588094005 @default.
• W2008478712 cites W1603869179 @default.
• W2008478712 cites W1606977653 @default.
• W2008478712 cites W1786370469 @default.
• W2008478712 cites W1873199501 @default.
• W2008478712 cites W1916639766 @default.
• W2008478712 cites W1986839556 @default.
• W2008478712 cites W2005912066 @default.
• W2008478712 cites W2006986935 @default.
• W2008478712 cites W2019054377 @default.
• W2008478712 cites W2021539568 @default.
• W2008478712 cites W2031645882 @default.
• W2008478712 cites W2059164909 @default.
• W2008478712 cites W2061700743 @default.
• W2008478712 cites W2070936740 @default.
• W2008478712 cites W2077700442 @default.
• W2008478712 cites W2085137948 @default.
• W2008478712 cites W2090256338 @default.
• W2008478712 cites W2100837269 @default.
• W2008478712 cites W4211027922 @default.
• W2008478712 cites W4231654880 @default.
• W2008478712 cites W4256299930 @default.
• W2008478712 cites W4293247451 @default.
• W2008478712 cites W4386008122 @default.
• W2008478712 doi "https://doi.org/10.1074/jbc.271.34.20690" @default.
• W2008478712 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/8702819" @default.
• W2008478712 hasPublicationYear "1996" @default.
• W2008478712 type Work @default.
• W2008478712 sameAs 2008478712 @default.
• W2008478712 citedByCount "69" @default.
• W2008478712 countsByYear W20084787122013 @default.
• W2008478712 countsByYear W20084787122014 @default.
• W2008478712 countsByYear W20084787122015 @default.
• W2008478712 countsByYear W20084787122016 @default.
• W2008478712 countsByYear W20084787122018 @default.
• W2008478712 countsByYear W20084787122021 @default.
• W2008478712 countsByYear W20084787122023 @default.
• W2008478712 crossrefType "journal-article" @default.
• W2008478712 hasAuthorship W2008478712A5005151768 @default.
• W2008478712 hasAuthorship W2008478712A5021046528 @default.
• W2008478712 hasBestOaLocation W20084787121 @default.
• W2008478712 hasConcept C104292427 @default.
• W2008478712 hasConcept C104317684 @default.
• W2008478712 hasConcept C106847996 @default.
• W2008478712 hasConcept C123894998 @default.
• W2008478712 hasConcept C153911025 @default.
• W2008478712 hasConcept C155672457 @default.
• W2008478712 hasConcept C171250308 @default.
• W2008478712 hasConcept C176406525 @default.
• W2008478712 hasConcept C178790620 @default.
• W2008478712 hasConcept C179247698 @default.
• W2008478712 hasConcept C181199279 @default.
• W2008478712 hasConcept C185592680 @default.
• W2008478712 hasConcept C192562407 @default.
• W2008478712 hasConcept C197404232 @default.
• W2008478712 hasConcept C204628709 @default.
• W2008478712 hasConcept C2776190903 @default.
• W2008478712 hasConcept C2778204606 @default.
• W2008478712 hasConcept C2778306172 @default.
• W2008478712 hasConcept C2780676925 @default.
• W2008478712 hasConcept C40767141 @default.
• W2008478712 hasConcept C55493867 @default.
• W2008478712 hasConcept C86803240 @default.
• W2008478712 hasConceptScore W2008478712C104292427 @default.
• W2008478712 hasConceptScore W2008478712C104317684 @default.
• W2008478712 hasConceptScore W2008478712C106847996 @default.
• W2008478712 hasConceptScore W2008478712C123894998 @default.
• W2008478712 hasConceptScore W2008478712C153911025 @default.
• W2008478712 hasConceptScore W2008478712C155672457 @default.
• W2008478712 hasConceptScore W2008478712C171250308 @default.
• W2008478712 hasConceptScore W2008478712C176406525 @default.
• W2008478712 hasConceptScore W2008478712C178790620 @default.
• W2008478712 hasConceptScore W2008478712C179247698 @default.
• W2008478712 hasConceptScore W2008478712C181199279 @default.
• W2008478712 hasConceptScore W2008478712C185592680 @default.

• next