Evidence for the involvement of the unique C-tail of S100A9 in the binding of arachidonic acid to the heterocomplex S100A8/A9.
Protein complexes formed by S100A8 and S100A9 represent the only AA-binding capacity in the human neutrophilic cytosol and are involved in the intracellular arachidonic acid metabolism. The formation of S100A8/A9 protein complexes and the binding of calcium to the complexes are prerequisites for the specific binding of polyunsaturated fatty acids. The present study was undertaken to characterize the fatty acid binding site within the protein complex. Deletions at both termini and point mutations of different basic amino acids especially within the extended C-terminal tail of human S100A9 were introduced. The S100A9 mutant proteins were then analyzed with respect to protein-protein interaction (GST pull down-assay and yeast two-hybrid system) and functional properties (arachidonic acid and calcium binding). The data give strong evidence that the unique C-tail of S100A9 containing the three consecutive histidine residues (His103-His105) represents the region to which the fatty acid carboxy-group is bound to the protein complex. The localization of the AA-binding site within the unique C-tail of S100A9 correlates with the fact that fatty acid binding has not yet been reported for other S100 proteins.
Sopalla
Claudia
C
Institute of Experimental Dermatology, University of Münster, von-Esmarch-Str. 58, D-48149 Münster, Germany.
Leukert
Nadja
N
Sorg
Clemens
C
Kerkhoff
Claus
C
D000595:Amino Acid Sequence; D024361:Amino Acids, Basic / Q000737:chemistry; D016718:Arachidonic Acid / Q000737:chemistry*; D002118:Calcium / Q000378:metabolism; D040501:Calgranulin A / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D017354:Point Mutation / Q000235:genetics; D011485:Protein Binding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017384:Sequence Deletion / Q000235:genetics; D014170:Transformation, Genetic
D024361:Amino Acids, Basic; D040501:Calgranulin A; D005227:Fatty Acids; D011994:Recombinant Proteins; D016718:Arachidonic Acid; D005982:Glutathione Transferase; D002118:Calcium