Detection and identification of protein interactions of S100 proteins by ProteinChip technology.
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.
Lehmann
Roland
R
Core Unit Chip Application (CUCA), Institute of Human Genetics and Anthropology, Friedrich-Schiller-University, 07740 Jena, Germany.
Melle
Christian
C
Escher
Niko
N
von Eggeling
Ferdinand
F
D017306:Annexin A2 / Q000737:chemistry; D002135:Calcium-Binding Proteins / Q000737:chemistry; D002460:Cell Line; D019992:Databases as Topic; D006801:Humans; D047468:Immunoprecipitation; D015603:Keratinocytes / Q000166:cytology; D010449:Peptide Mapping; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D000075023:S100 Calcium Binding Protein A7; D009418:S100 Proteins / Q000032:analysis* / Q000737:chemistry*; D020539:Sequence Analysis, Protein; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000494:pharmacology; D000099248:S100 Calcium Binding Protein A10
D017306:Annexin A2; D002135:Calcium-Binding Proteins; D020543:Proteome; D000075023:S100 Calcium Binding Protein A7; D009418:S100 Proteins; D014357:Trypsin; D000099248:S100 Calcium Binding Protein A10; C089242:S100A7 protein, human