Self-assembling protein microarrays

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

Research output: Contribution to journalArticle

458 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)86-90
Number of pages5
JournalScience
Volume305
Issue number5680
DOIs
StatePublished - Jul 2 2004
Externally publishedYes

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Protein Array Analysis
Proteins
Geminin
Printing
Protein Stability
Reticulocytes
DNA Replication
Glass
Epitopes
Complementary DNA
Technology

ASJC Scopus subject areas

  • General

Cite this

Ramachandran, N., Hainsworth, E., Bhullar, B., Eisenstein, S., Rosen, B., Lau, A. Y., ... LaBaer, J. (2004). Self-assembling protein microarrays. Science, 305(5680), 86-90. https://doi.org/10.1126/science.1097639

Self-assembling protein microarrays. / Ramachandran, Niroshan; Hainsworth, Eugenie; Bhullar, Bhupinder; Eisenstein, Samuel; Rosen, Benjamin; Lau, Albert Y.; Walter, Johannes C.; LaBaer, Joshua.

In: Science, Vol. 305, No. 5680, 02.07.2004, p. 86-90.

Research output: Contribution to journalArticle

Ramachandran, N, Hainsworth, E, Bhullar, B, Eisenstein, S, Rosen, B, Lau, AY, Walter, JC & LaBaer, J 2004, 'Self-assembling protein microarrays', Science, vol. 305, no. 5680, pp. 86-90. https://doi.org/10.1126/science.1097639
Ramachandran N, Hainsworth E, Bhullar B, Eisenstein S, Rosen B, Lau AY et al. Self-assembling protein microarrays. Science. 2004 Jul 2;305(5680):86-90. https://doi.org/10.1126/science.1097639
Ramachandran, Niroshan ; Hainsworth, Eugenie ; Bhullar, Bhupinder ; Eisenstein, Samuel ; Rosen, Benjamin ; Lau, Albert Y. ; Walter, Johannes C. ; LaBaer, Joshua. / Self-assembling protein microarrays. In: Science. 2004 ; Vol. 305, No. 5680. pp. 86-90.
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