Profiling of generic anti-phosphopeptide antibodies and kinases with peptide microarrays using radioactive and fluorescence-based assays

Molecular Diversity
Sören PanseJens Schneider-Mergener

Abstract

Kinases represent one of the largest enzyme families and key regulatory proteins in the cell. Only a small subset of these enzymes has been characterised so far. We have prepared different types of phosphopeptide and peptide microarrays displaying peptides deduced from annotated human phosphorylation sites and cytoplasmic domains of all annotated human membrane proteins. This approach was enabled by fully-automated high throughput micro-scale synthesis of peptides by the SPOT technology combined with chemo-selective immobilisation on modified glass slides. The phosphopeptide microarrays displaying 2923 peptides in total have been used for the characterisation of commercially available generic anti-phosphopeptide antibodies. This enabled us to detect Abl kinase activity on a microarray with anti-phosphotyrosine antibodies yielding results comparable to those obtained from a radioactive assay. More than 13 000 peptides deposited on six glass slides were used to profile casein kinase 2 (CK2) using a radioactive assay, since no generic antibody for the reliable detection of serine or threonine phosphorylation could be identified. All previously identified substrates were detected in the microarray experiment. In order to confirm wh...Continue Reading

Citations

Mar 26, 2005·Chembiochem : a European Journal of Chemical Biology·Kin-ya TomizakiHisakazu Mihara
Jan 12, 2007·Molecular BioSystems·Johann Grognux, Jean-Louis Reymond
Dec 22, 2010·Biointerphases·M Uttamchandani, S Moochhala
Jan 21, 2010·Assay and Drug Development Technologies·Kenji TanakaHideya Komatani
Sep 13, 2014·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·K OtsuS C Dreskin
Mar 3, 2005·Chembiochem : a European Journal of Chemical Biology·Mike SchutkowskiUlf Reimer
Dec 7, 2007·Biotechnology & Genetic Engineering Reviews·Kai HilperRobert E W Hancock
Oct 22, 2010·Epigenetics : Official Journal of the DNA Methylation Society·Ina BockAlbert Jeltsch
May 24, 2011·Molecular Biotechnology·Alexandra ThieleMike Schutkowski
Aug 2, 2013·Avian Diseases·Dusan Kunec
Jul 18, 2019·MAbs·Nileena VelappanAndrew Bradbury
Oct 6, 2005·Journal of Biochemistry and Molecular Biology·Dong-Sik ShinYoon-Sik Lee
Jan 9, 2008·Analytical Chemistry·Kazuki InamoriYoshiki Katayama
Dec 15, 2005·Analytical Chemistry·Nicholas RupcichJohn D Brennan

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