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PubMed · 15232106

Self-assembling protein microarrays.

Niroshan Ramachandran·Eugenie Hainsworth·Bhupinder Bhullar·Samuel Eisenstein·Benjamin Rosen·Albert Y Lau·Johannes C Walter·Joshua LaBaer
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Abstract

Protein microarrays provide a powerful tool for the study of protein function. However, they are not widely used, in part because of the challenges in producing proteins to spot on the arrays. We generated protein microarrays by printing complementary DNAs onto glass slides and then translating target proteins with mammalian reticulocyte lysate. Epitope tags fused to the proteins allowed them to be immobilized in situ. This obviated the need to purify proteins, avoided protein stability problems during storage, and captured sufficient protein for functional studies. We used the technology to map pairwise interactions among 29 human DNA replication initiation proteins, recapitulate the regulation of Cdt1 binding to select replication proteins, and map its geminin-binding domain.

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Cell Cycle ProteinsCell-Free SystemDNA ReplicationDNA, ComplementaryEpitopesGemininHumansMinichromosome Maintenance Complex Component 2Minichromosome Maintenance Complex Component 6Nuclear ProteinsProtein Array AnalysisProtein BindingProtein BiosynthesisProtein Interaction MappingProtein Structure, TertiaryProteinsReplication OriginTranscription, GeneticJournal ArticleResearch Support, U.S. Gov't, P.H.S.

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Niroshan Ramachandran, Eugenie Hainsworth, Bhupinder Bhullar, Samuel Eisenstein, Benjamin Rosen, Albert Y Lau, Johannes C Walter, Joshua LaBaer. 2004-07-02. Self-assembling protein microarrays.. https://doi.org/10.1126/science.1097639

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