Multiple chromatographic forms of the formylpeptide chemoattractant receptor and their relationship to GTP-binding proteins.
The radiolabeled formylpeptide chemoattractant receptor, partially purified by wheat germ agglutinin-Sepharose chromatography, eluted in three distinct peaks when chromatographed on DEAE-Fractogel. Incubation of the lectin-Sepharose purified receptor with 100 microM guanosine 5-(3-O-thio) triphosphate markedly altered the distribution of the radiolabeled receptor when chromatographed on the ion exchange resin. Peaks 2 and 3 were reduced by approximately 50% while peak 1 was concomitantly increased. Western blot analysis revealed the presence of a 40 kDa GTP-binding protein alpha-subunit only in peak 3. Incubation of Western blots with [alpha-32P]GTP detected low molecular mass GTP-binding proteins (24 and 26 kDa) that coeluted with the receptor in peak 2. Incubation of the peak 1 receptor fractions with fractions containing a mixture of GTP binding proteins resulted in the generation of peaks 2 and 3 when chromatographed on DEAE-Fractogel. These results demonstrate that the chromatographic behavior of the formylpeptide receptor is dependent upon its association with GTP binding proteins and that more than one type of GTP-binding protein may be involved.
Polakis
P G
PG
Department of Pharmacological Sciences, Genentech, Inc., South San Francisco, CA 94080.
Evans
T
T
Snyderman
R
R
D002460:Cell Line; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D019204:GTP-Binding Proteins / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000378:metabolism*; D011869:Radioligand Assay; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic / Q000032:analysis*
D008565:Membrane Proteins; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic; D009240:N-Formylmethionine Leucyl-Phenylalanine; D019204:GTP-Binding Proteins