The Rh polypeptide is a major fatty acid-acylated erythrocyte membrane protein

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

The erythrocyte Rh antigens contain an M(r) = 32,000 integral protein which is thought to contribute in some way to the organization of surrounding phospholipid. To search for possible fatty acid acylation of the Rh polypeptide, intact human erythrocytes were incubated with [3H]palmitic acid prior to preparation of membranes and sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. Several membrane proteins were labeled, but none corresponded to the glycophorins or membrane proteins 1-8. An M(r) = 32,000 band was prominently labeled on Rh (D)-negative and -positive erythrocytes and could be precipitated from the latter with anti-D. No similar protein was labeled on membranes from Rh(mod) erythrocytes, a rare phenotype lacking Rh antigens. Labeling of the Rh polypeptide most likely represents palmitic acid acylation through thioester linkages. The 3H label was not extracted with chloroform/methanol, but was quantitatively eluted with hydroxylamine and co-chromatographed with palmitohydroxamate and free palmitate by thin layer chromatography. The fatty acid acylations occurred independent of protein synthesis and were completely reversed by chase with unlabeled palmitate. It is concluded that the Rh polypeptide is fatty acid-acylated, being a major substrate of an acylation-deacylation mechanism associated with the erythrocyte membrane.

Original languageEnglish (US)
Pages (from-to)18193-18196
Number of pages4
JournalJournal of Biological Chemistry
Volume263
Issue number34
StatePublished - Dec 1 1988

Fingerprint

Acylation
Erythrocyte Membrane
Membrane Proteins
Fatty Acids
Erythrocytes
Peptides
Palmitic Acid
Palmitates
Membranes
Photofluorography
Glycophorin
Antigens
Thin layer chromatography
Hydroxylamine
Proteins
Thin Layer Chromatography
Chloroform
Electrophoresis
Sodium Dodecyl Sulfate
Labeling

ASJC Scopus subject areas

  • Biochemistry

Cite this

The Rh polypeptide is a major fatty acid-acylated erythrocyte membrane protein. / DeVetten, Marcel P; Agre, P.

In: Journal of Biological Chemistry, Vol. 263, No. 34, 01.12.1988, p. 18193-18196.

Research output: Contribution to journalArticle

@article{17284c01299745aaa501f223d8978fd8,
title = "The Rh polypeptide is a major fatty acid-acylated erythrocyte membrane protein",
abstract = "The erythrocyte Rh antigens contain an M(r) = 32,000 integral protein which is thought to contribute in some way to the organization of surrounding phospholipid. To search for possible fatty acid acylation of the Rh polypeptide, intact human erythrocytes were incubated with [3H]palmitic acid prior to preparation of membranes and sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. Several membrane proteins were labeled, but none corresponded to the glycophorins or membrane proteins 1-8. An M(r) = 32,000 band was prominently labeled on Rh (D)-negative and -positive erythrocytes and could be precipitated from the latter with anti-D. No similar protein was labeled on membranes from Rh(mod) erythrocytes, a rare phenotype lacking Rh antigens. Labeling of the Rh polypeptide most likely represents palmitic acid acylation through thioester linkages. The 3H label was not extracted with chloroform/methanol, but was quantitatively eluted with hydroxylamine and co-chromatographed with palmitohydroxamate and free palmitate by thin layer chromatography. The fatty acid acylations occurred independent of protein synthesis and were completely reversed by chase with unlabeled palmitate. It is concluded that the Rh polypeptide is fatty acid-acylated, being a major substrate of an acylation-deacylation mechanism associated with the erythrocyte membrane.",
author = "DeVetten, {Marcel P} and P. Agre",
year = "1988",
month = "12",
day = "1",
language = "English (US)",
volume = "263",
pages = "18193--18196",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "34",

}

TY - JOUR

T1 - The Rh polypeptide is a major fatty acid-acylated erythrocyte membrane protein

AU - DeVetten, Marcel P

AU - Agre, P.

PY - 1988/12/1

Y1 - 1988/12/1

N2 - The erythrocyte Rh antigens contain an M(r) = 32,000 integral protein which is thought to contribute in some way to the organization of surrounding phospholipid. To search for possible fatty acid acylation of the Rh polypeptide, intact human erythrocytes were incubated with [3H]palmitic acid prior to preparation of membranes and sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. Several membrane proteins were labeled, but none corresponded to the glycophorins or membrane proteins 1-8. An M(r) = 32,000 band was prominently labeled on Rh (D)-negative and -positive erythrocytes and could be precipitated from the latter with anti-D. No similar protein was labeled on membranes from Rh(mod) erythrocytes, a rare phenotype lacking Rh antigens. Labeling of the Rh polypeptide most likely represents palmitic acid acylation through thioester linkages. The 3H label was not extracted with chloroform/methanol, but was quantitatively eluted with hydroxylamine and co-chromatographed with palmitohydroxamate and free palmitate by thin layer chromatography. The fatty acid acylations occurred independent of protein synthesis and were completely reversed by chase with unlabeled palmitate. It is concluded that the Rh polypeptide is fatty acid-acylated, being a major substrate of an acylation-deacylation mechanism associated with the erythrocyte membrane.

AB - The erythrocyte Rh antigens contain an M(r) = 32,000 integral protein which is thought to contribute in some way to the organization of surrounding phospholipid. To search for possible fatty acid acylation of the Rh polypeptide, intact human erythrocytes were incubated with [3H]palmitic acid prior to preparation of membranes and sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. Several membrane proteins were labeled, but none corresponded to the glycophorins or membrane proteins 1-8. An M(r) = 32,000 band was prominently labeled on Rh (D)-negative and -positive erythrocytes and could be precipitated from the latter with anti-D. No similar protein was labeled on membranes from Rh(mod) erythrocytes, a rare phenotype lacking Rh antigens. Labeling of the Rh polypeptide most likely represents palmitic acid acylation through thioester linkages. The 3H label was not extracted with chloroform/methanol, but was quantitatively eluted with hydroxylamine and co-chromatographed with palmitohydroxamate and free palmitate by thin layer chromatography. The fatty acid acylations occurred independent of protein synthesis and were completely reversed by chase with unlabeled palmitate. It is concluded that the Rh polypeptide is fatty acid-acylated, being a major substrate of an acylation-deacylation mechanism associated with the erythrocyte membrane.

UR - http://www.scopus.com/inward/record.url?scp=0024203554&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024203554&partnerID=8YFLogxK

M3 - Article

VL - 263

SP - 18193

EP - 18196

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 34

ER -