Lipid-binding Activity of Intrinsically Unstructured Cytoplasmic Domains of Multichain Immune Recognition Receptor Signaling Subunits
Cell Signaling, Lipids, Peptides and Proteins, Receptors, Vesicles
Digital Object Identifier (DOI)
Multichain immune recognition receptors (MIRRs) found on the surface of T cells, B cells, mast cells, natural killer cells, basophils, and other immune cells are formed by the association of several single-pass transmembrane proteins, with immunoglobulin-like ligand recognition domains and signal-transducing domains present on separate subunits. The MIRR signaling subunits all have cytoplasmic domains containing one or more copies of an immunoreceptor tyrosine-based activation motif (ITAM), tyrosine residues of which are phosphorylated upon receptor engagement in an early and obligatory event in the signaling cascade. Despite the proximity to the cell membrane and crucial role in transmembrane signal transduction, little is known about the structure and lipid-binding activity of the ITAM-containing cytoplasmic domains. Here we investigate the conformation and lipid-binding activity of several MIRR cytoplasmic domains, namely, T cell receptor ζcyt, CD3εcyt, CD3δcyt, and CD3γcyt, B cell receptor Igαcyt and Igβcyt, and Fc receptor FcεRIγcyt, using purified recombinant proteins. Secondary structure prediction analysis and experimental circular dichroism spectra identify each of these cytoplasmic domains as natively unfolded proteins. We also report that ζcyt, CD3εcyt, and FcεRIγcyt bind to acidic and mixed phospholipid vesicles and that the binding strength correlates with the protein net charge and the presence of clustered basic amino acid residues. Circular dichroism analysis reveals the lack of secondary structure for these domains in lipid-bound form. Phosphorylation of ζcyt and FcεRIγcyt does not alter their random-coil conformation but weakens binding to membranes. The implications of these results for transmembrane signal transduction by immune receptors are discussed.
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Citation / Publisher Attribution
Biochemistry, v. 45, issue 51, p. 14731-15739
Scholar Commons Citation
Sigalov, Alexander B.; Aivazian, Dikran A.; Uversky, Vladimir N.; and Stern, Lawrence J., "Lipid-binding Activity of Intrinsically Unstructured Cytoplasmic Domains of Multichain Immune Recognition Receptor Signaling Subunits" (2006). Molecular Medicine Faculty Publications. 770.