Detection and identification of protein interactions of S100 proteins by ProteinChip technology

Roland Lehmann; Christian Melle; Niko Escher; Ferdinand von Eggeling

doi:10.1021/pr050163s

Detection and identification of protein interactions of S100 proteins by ProteinChip technology

J Proteome Res. 2005 Sep-Oct;4(5):1717-21. doi: 10.1021/pr050163s.

Authors

Roland Lehmann¹, Christian Melle, Niko Escher, Ferdinand von Eggeling

Affiliation

¹ Core Unit Chip Application (CUCA), Institute of Human Genetics and Anthropology, Friedrich-Schiller-University, 07740 Jena, Germany.

PMID: 16212425
DOI: 10.1021/pr050163s

Abstract

The aim of this work was to establish an approach for identification of protein interactions. This assay used an anti-S100A8 antibody coupled on beads and incubated with cell extract. The bead eluates were analyzed using ProteinChip technology and subsequently subjected to an appropriate digestion. Molecular masses of digestion fragments were determined by SELDI-MS, and database analysis revealed S100A10 as interacting protein. This result was confirmed by co-immunoprecipitation and immunocapturing. Using S100A10 as new bait, a specific interaction with S100A7 was detectable.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Annexin A2 / chemistry
Calcium-Binding Proteins / chemistry
Cell Line
Databases as Topic
Humans
Immunoprecipitation
Keratinocytes / cytology
Peptide Mapping
Protein Array Analysis / methods*
Protein Binding
Proteome*
Proteomics / methods*
S100 Calcium Binding Protein A7
S100 Proteins / analysis*
S100 Proteins / chemistry*
Sequence Analysis, Protein
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Trypsin / pharmacology

Substances

Annexin A2
Calcium-Binding Proteins
Proteome
S100 Calcium Binding Protein A7
S100 Proteins
S100 calcium binding protein A10
S100A7 protein, human
Trypsin