Journal
PROTEOMICS
Volume 5, Issue 2, Pages 416-419Publisher
WILEY
DOI: 10.1002/pmic.200400948
Keywords
immobilized metal ion affinity chromatography; protein microarray; protein orientation; sulfotransferase
Funding
- NCRR NIH HHS [1R43RR17130-01] Funding Source: Medline
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We compare the catalytic activities of enzymes immobilized on silicon surfaces with and without orientation. While oriented sulfotransferases selectively immobilized on an otherwise zero-background surface via 6XHis tags faithfully reflect activities of solution phase enzymes, those with random orientation on the surface do not. This finding demonstrates that controlling the orientation of immobilized protein molecules and designing an ideal local chemical environment on the solid surface are both essential if protein microarrays are to be used as quantitative tools in biomedical research.
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