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Science 2 July 2004:
Vol. 305. no. 5680, pp. 86 - 90
DOI: 10.1126/science.1097639
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Reports

Self-Assembling Protein Microarrays

Niroshan Ramachandran,1 Eugenie Hainsworth,1,2 Bhupinder Bhullar,1 Samuel Eisenstein,1 Benjamin Rosen,1 Albert Y. Lau,1 Johannes C. Walter,3 Joshua LaBaer1*

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.

1 Harvard Institute of Proteomics, Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 320 Charles Street, Cambridge, MA 02141, USA.
2 Technology and Engineering Center, Harvard Medical School, 240 Long-wood Avenue, Boston, MA 02115, USA.
3 Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Long-wood Avenue, Boston, MA 02115, USA.

* To whom correspondence should be addressed. E-mail: josh{at}hms.harvard.edu

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Science. ISSN 0036-8075 (print), 1095-9203 (online)