Identification and purification of a novel G protein from neutrophils

Burton F. Dickey; Hae Yung Pyun; Kim C. Williamson; Javier Navarro

doi:10.1016/0014-5793(87)80237-5

Identification and purification of a novel G protein from neutrophils

Burton F. Dickey, Hae Yung Pyun, Kim C. Williamson, Javier Navarro

Research output: Contribution to journal › Article › peer-review

15 Scopus citations

Abstract

A novel G protein which appears to couple chemotactic peptide receptors to a polyphosphoinositide phospholipase C has been purified from rabbit neutrophils. Neutrophil membranes were solubilized with sodium cholate and fractionated by successive anion exchange, gel filtration and hydrophobic chromatography. Guanosine-5′-(3-O-thio)triphosphate binding activity was purified 170-fold from the soluble extract. The α-subunit of the purified G protein was identified by pertussis toxin-catalyzed ADP-ribosylation, and found to have an M_r of 40 000. The β-subunit (M_r 36 000) comigrated on SDS-polyacrylamide gel electrophoresis with the β-subunits of bovine brain Gi and Go. The neutrophil pertussis toxin substrate is highly unstable in cholate solution unless 30% ethylene glycol is added. Structural and functional analysis of this novel G protein will advance our understanding of the molecular mechanisms of coupling of receptors to phospholipase C.

Original language	English (US)
Pages (from-to)	289-292
Number of pages	4
Journal	FEBS Letters
Volume	219
Issue number	2
DOIs	https://doi.org/10.1016/0014-5793(87)80237-5
State	Published - Jul 27 1987
Externally published	Yes

Keywords

(Rabbit neutrophil)
Guanine nucleotide-binding protein
Phospholipase C
fMet-Leu-Phe receptor

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1016/0014-5793(87)80237-5

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title = "Identification and purification of a novel G protein from neutrophils",

abstract = "A novel G protein which appears to couple chemotactic peptide receptors to a polyphosphoinositide phospholipase C has been purified from rabbit neutrophils. Neutrophil membranes were solubilized with sodium cholate and fractionated by successive anion exchange, gel filtration and hydrophobic chromatography. Guanosine-5′-(3-O-thio)triphosphate binding activity was purified 170-fold from the soluble extract. The α-subunit of the purified G protein was identified by pertussis toxin-catalyzed ADP-ribosylation, and found to have an Mr of 40 000. The β-subunit (Mr 36 000) comigrated on SDS-polyacrylamide gel electrophoresis with the β-subunits of bovine brain Gi and Go. The neutrophil pertussis toxin substrate is highly unstable in cholate solution unless 30% ethylene glycol is added. Structural and functional analysis of this novel G protein will advance our understanding of the molecular mechanisms of coupling of receptors to phospholipase C.",

keywords = "(Rabbit neutrophil), Guanine nucleotide-binding protein, Phospholipase C, fMet-Leu-Phe receptor",

author = "Dickey, {Burton F.} and Pyun, {Hae Yung} and Williamson, {Kim C.} and Javier Navarro",

note = "Funding Information: This work was supported by grants SK1 lHLO55 030-02 to B.F.D. from USPHS and DCB-8511671 to J.N. from NSF.",

year = "1987",

month = jul,

day = "27",

doi = "10.1016/0014-5793(87)80237-5",

language = "English (US)",

volume = "219",

pages = "289--292",

journal = "FEBS Letters",

issn = "0014-5793",

publisher = "John Wiley and Sons Inc.",

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T1 - Identification and purification of a novel G protein from neutrophils

AU - Dickey, Burton F.

AU - Pyun, Hae Yung

AU - Williamson, Kim C.

AU - Navarro, Javier

N1 - Funding Information: This work was supported by grants SK1 lHLO55 030-02 to B.F.D. from USPHS and DCB-8511671 to J.N. from NSF.

PY - 1987/7/27

Y1 - 1987/7/27

N2 - A novel G protein which appears to couple chemotactic peptide receptors to a polyphosphoinositide phospholipase C has been purified from rabbit neutrophils. Neutrophil membranes were solubilized with sodium cholate and fractionated by successive anion exchange, gel filtration and hydrophobic chromatography. Guanosine-5′-(3-O-thio)triphosphate binding activity was purified 170-fold from the soluble extract. The α-subunit of the purified G protein was identified by pertussis toxin-catalyzed ADP-ribosylation, and found to have an Mr of 40 000. The β-subunit (Mr 36 000) comigrated on SDS-polyacrylamide gel electrophoresis with the β-subunits of bovine brain Gi and Go. The neutrophil pertussis toxin substrate is highly unstable in cholate solution unless 30% ethylene glycol is added. Structural and functional analysis of this novel G protein will advance our understanding of the molecular mechanisms of coupling of receptors to phospholipase C.

AB - A novel G protein which appears to couple chemotactic peptide receptors to a polyphosphoinositide phospholipase C has been purified from rabbit neutrophils. Neutrophil membranes were solubilized with sodium cholate and fractionated by successive anion exchange, gel filtration and hydrophobic chromatography. Guanosine-5′-(3-O-thio)triphosphate binding activity was purified 170-fold from the soluble extract. The α-subunit of the purified G protein was identified by pertussis toxin-catalyzed ADP-ribosylation, and found to have an Mr of 40 000. The β-subunit (Mr 36 000) comigrated on SDS-polyacrylamide gel electrophoresis with the β-subunits of bovine brain Gi and Go. The neutrophil pertussis toxin substrate is highly unstable in cholate solution unless 30% ethylene glycol is added. Structural and functional analysis of this novel G protein will advance our understanding of the molecular mechanisms of coupling of receptors to phospholipase C.

KW - (Rabbit neutrophil)

KW - Guanine nucleotide-binding protein

KW - Phospholipase C

KW - fMet-Leu-Phe receptor

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DO - 10.1016/0014-5793(87)80237-5

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JO - FEBS Letters

JF - FEBS Letters

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ER -

Identification and purification of a novel G protein from neutrophils

Abstract

Keywords

ASJC Scopus subject areas

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