Characterization of complex formation between Gi2 and octyl glucoside solubilized neutrophil N-formyl peptide chemoattractant receptor by sedimentation velocity

International Union of Basic and Clinical Pharmacology. LXXIII. Nomenclature for the Formyl Peptide Receptor (FPR) Family

“…Second, G␣ 16 transcript abundance is very low in mature neutrophils, and induction of HL-60 cell differentiation is associated with a progressive decrease in expression of G␣ 16 and an increase in the expression of G i ␣ 2 (Amatruda et al, 1991). Third, a large body of evidence indicates that the fMLF-induced responses require physical association of FPR1 with PTX-sensitive G i ␣ 2 protein in leukocytes (Bommakanti et al, 1992(Bommakanti et al, , 1993(Bommakanti et al, , 1994(Bommakanti et al, , 1995Schreiber et al, 1993). However, residual molecules of G␣ 16 may account for a small PTX-resistant activity that has been observed in neutrophils, monocytes, and differentiated HL-60 cells stimulated with fMLF (Verghese et al, 1987).…”

“…The latter approach is based on the use of synthetic receptor-mimetic peptides to block the receptor-G protein interactions. A competition assay based on the sedimentation of a GTP-sensitive FPR1-G␣ i2 complex on sucrose gradients in the presence of octyl glucoside was developed by Bommakanti et al (1992Bommakanti et al ( , 1994 to assess the effect of FPR1-derived peptides on the conversion of FPR1 from a 7S (FPR1-G i complex) to a 4S species (FPR1 only). Synthetic FPR1-derived peptides were also used by Schreiber et al (1994) in a competitive enzymelinked immunosorbent assay.…”

Extensive Contact between Gi2 and N-Formyl Peptide Receptor of Human Neutrophils: Mapping of Binding Sites Using Receptor-Mimetic Peptides

“…The 7S form can be produced from 4S FPR by adding Gl2, and the reconstituted 7S is quantitatively dissociated to 4S by guanine nucleotides (Bommakanti et al, 1992) and is immunosedimentable by anti-Gi2a antibodies (Bommakanti et al, 1992). The 7S form of FPR is a functional, physical complex with the Gj2 protein since it contains agonist occupied receptor and an empty guanine nucleotide binding site (Bommakanti et al, 1994) required for receptor-mediated GDP release and GTP exchange.…”

Real-time Analysis of G Protein-coupled Receptor Reconstitution in a Solubilized System

“…Comparable efforts are being made to describe the molecular mechanisms that influence the specificity of coupling interactions between GPCR and their cognate heterotrimeric G proteins. Several experimental systems for reconstitution provide alternatives for studying receptors and G proteins, such as adding G protein to stripped membranes, pairing receptors and G proteins in phospholipid vesicles (3), and examining solubilized receptor-G protein interactions using gradient centrifugation (4).…”