Membrane orientation of Rh(D) polypeptide and partial localization of its epitope-containing domain

Kimita Suyama, Jack Goldstein

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

We have previously shown that the effects of various enzyme treatments on Rh antigen-containing polypeptides in situ could be monitored by an antibody preparation which recognizes only these polypeptides following Western blotting. We now have prepared antibodies that specifically react with either the N- or C-terminal ends of Rh-related proteins. Using all three, we have established that the C-terminus of Rh(D) polypeptide is at the cell surface, whereas its N-terminal domain is situated at the cytoplasmic side of the red blood cell membrane. Chymotrypsin digestion of ghosts derived from (-D-/-D-) cells that are devoid of Rh (C/c) and (E/e) antigens produces three major Rh(D)-related fragments: the 20-Kd fragment contains the molecule's C-terminal domain, the 17-Kd fragment its N-terminus, and the 13-Kd fragment neither. However, only the 17-Kd fragment forms an immune-complex with human polyclonal anti-D, indicating that it contains the Rh(D) antigenic domain. Other findings presented here provide further evidence for a unique folding of Rh(D) polypeptide within the cell membrane and suggest that Rh(CXc) and (E/e) polypeptides, when present, may form complexes with it.

Original languageEnglish (US)
Pages (from-to)808-812
Number of pages5
JournalBlood
Volume79
Issue number3
StatePublished - Feb 1 1992
Externally publishedYes

Fingerprint

Epitopes
Membranes
Peptides
Cell membranes
Cell Membrane
Hepatitis B e Antigens
Somatostatin-Secreting Cells
Antibodies
Chymotrypsin
Antigen-Antibody Complex
Digestion
Blood
Erythrocytes
Western Blotting
Antigens
Molecules
Enzymes
Proteins

ASJC Scopus subject areas

  • Hematology

Cite this

Membrane orientation of Rh(D) polypeptide and partial localization of its epitope-containing domain. / Suyama, Kimita; Goldstein, Jack.

In: Blood, Vol. 79, No. 3, 01.02.1992, p. 808-812.

Research output: Contribution to journalArticle

@article{ee7dad4f03d14c7291afc10642457b98,
title = "Membrane orientation of Rh(D) polypeptide and partial localization of its epitope-containing domain",
abstract = "We have previously shown that the effects of various enzyme treatments on Rh antigen-containing polypeptides in situ could be monitored by an antibody preparation which recognizes only these polypeptides following Western blotting. We now have prepared antibodies that specifically react with either the N- or C-terminal ends of Rh-related proteins. Using all three, we have established that the C-terminus of Rh(D) polypeptide is at the cell surface, whereas its N-terminal domain is situated at the cytoplasmic side of the red blood cell membrane. Chymotrypsin digestion of ghosts derived from (-D-/-D-) cells that are devoid of Rh (C/c) and (E/e) antigens produces three major Rh(D)-related fragments: the 20-Kd fragment contains the molecule's C-terminal domain, the 17-Kd fragment its N-terminus, and the 13-Kd fragment neither. However, only the 17-Kd fragment forms an immune-complex with human polyclonal anti-D, indicating that it contains the Rh(D) antigenic domain. Other findings presented here provide further evidence for a unique folding of Rh(D) polypeptide within the cell membrane and suggest that Rh(CXc) and (E/e) polypeptides, when present, may form complexes with it.",
author = "Kimita Suyama and Jack Goldstein",
year = "1992",
month = "2",
day = "1",
language = "English (US)",
volume = "79",
pages = "808--812",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "3",

}

TY - JOUR

T1 - Membrane orientation of Rh(D) polypeptide and partial localization of its epitope-containing domain

AU - Suyama, Kimita

AU - Goldstein, Jack

PY - 1992/2/1

Y1 - 1992/2/1

N2 - We have previously shown that the effects of various enzyme treatments on Rh antigen-containing polypeptides in situ could be monitored by an antibody preparation which recognizes only these polypeptides following Western blotting. We now have prepared antibodies that specifically react with either the N- or C-terminal ends of Rh-related proteins. Using all three, we have established that the C-terminus of Rh(D) polypeptide is at the cell surface, whereas its N-terminal domain is situated at the cytoplasmic side of the red blood cell membrane. Chymotrypsin digestion of ghosts derived from (-D-/-D-) cells that are devoid of Rh (C/c) and (E/e) antigens produces three major Rh(D)-related fragments: the 20-Kd fragment contains the molecule's C-terminal domain, the 17-Kd fragment its N-terminus, and the 13-Kd fragment neither. However, only the 17-Kd fragment forms an immune-complex with human polyclonal anti-D, indicating that it contains the Rh(D) antigenic domain. Other findings presented here provide further evidence for a unique folding of Rh(D) polypeptide within the cell membrane and suggest that Rh(CXc) and (E/e) polypeptides, when present, may form complexes with it.

AB - We have previously shown that the effects of various enzyme treatments on Rh antigen-containing polypeptides in situ could be monitored by an antibody preparation which recognizes only these polypeptides following Western blotting. We now have prepared antibodies that specifically react with either the N- or C-terminal ends of Rh-related proteins. Using all three, we have established that the C-terminus of Rh(D) polypeptide is at the cell surface, whereas its N-terminal domain is situated at the cytoplasmic side of the red blood cell membrane. Chymotrypsin digestion of ghosts derived from (-D-/-D-) cells that are devoid of Rh (C/c) and (E/e) antigens produces three major Rh(D)-related fragments: the 20-Kd fragment contains the molecule's C-terminal domain, the 17-Kd fragment its N-terminus, and the 13-Kd fragment neither. However, only the 17-Kd fragment forms an immune-complex with human polyclonal anti-D, indicating that it contains the Rh(D) antigenic domain. Other findings presented here provide further evidence for a unique folding of Rh(D) polypeptide within the cell membrane and suggest that Rh(CXc) and (E/e) polypeptides, when present, may form complexes with it.

UR - http://www.scopus.com/inward/record.url?scp=0026543111&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026543111&partnerID=8YFLogxK

M3 - Article

VL - 79

SP - 808

EP - 812

JO - Blood

JF - Blood

SN - 0006-4971

IS - 3

ER -