Affinity purification of an archaeal DNA replication protein network.

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TitleAffinity purification of an archaeal DNA replication protein network.
Publication TypeJournal Article
Year of Publication2010
AuthorsLi, Z, Santangelo, TJ, Cuboňová, L'ubomíra, Reeve, JN, Kelman, Z
Date Published2010
KeywordsArchaeal Proteins, Chromatography, Affinity, Complex Mixtures, DNA Replication, DNA, Archaeal, Mass Spectrometry, Protein Binding, Protein Interaction Mapping, Recombinant Proteins, Thermococcus

Nineteen Thermococcus kodakarensis strains have been constructed, each of which synthesizes a different His(6)-tagged protein known or predicted to be a component of the archaeal DNA replication machinery. Using the His(6)-tagged proteins, stable complexes assembled in vivo have been isolated directly from clarified cell lysates and the T. kodakarensis proteins present have been identified by mass spectrometry. Based on the results obtained, a network of interactions among the archaeal replication proteins has been established that confirms previously documented and predicted interactions, provides experimental evidence for previously unrecognized interactions between proteins with known functions and with unknown functions, and establishes a firm experimental foundation for archaeal replication research. The proteins identified and their participation in archaeal DNA replication are discussed and related to their bacterial and eukaryotic counterparts.

Alternate JournalMBio
PubMed ID20978540
PubMed Central IDPMC2962436
Grant List1F32-GM073336 / GM / NIGMS NIH HHS / United States
R01 GM053185 / GM / NIGMS NIH HHS / United States
R01GM53185 / GM / NIGMS NIH HHS / United States