In vitro incorporation of GPI-anchored proteins into human erythrocytes and their fate in the membrane

被引:47
作者
Civenni, G
Test, ST
Brodbeck, U
Bütikofer, P
机构
[1] Univ Bern, Inst Biochem & Mol Biol, CH-3012 Bern, Switzerland
[2] Childrens Hosp Oakland, Res Inst, Oakland, CA 94609 USA
关键词
D O I
10.1182/blood.V91.5.1784.1784_1784_1792
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
In many different cells, glycosylphosphatidylinositol (GPI)-anchored molecules are clustered in membrane microdomains that resist extraction by detergents at 4 degrees C. In this report, we identified the presence of such domains in human erythrocytes and examined the ability of exogenously-added GPI-anchored molecules to colocalize with the endogenous GPI-anchored proteins in these detergent-insoluble complexes. We found that the addition to human erythrocytes of three purified GPI-anchored proteins having different GPI lipid moieties resulted in their efficient and correct incorporation into the membrane. The extent of membrane insertion was dependent on the intactness of the GPI lipid moiety. However, unlike the endogenous GPI-anchored proteins, the in vitro incorporated GPI molecules were not resistant to membrane extraction by Triton X-100 at 4 degrees C. In addition, in contrast to the endogenous GPI-anchored proteins, they were not preferentially released from erythrocytes during vesiculation induced by calcium loading of the cells, These results suggest that in vitro incorporated GPI-linked molecules are excluded from pre-existing GPI-enriched membrane areas in human erythrocytes and that these microdomains may represent the sites of membrane vesicle formation. (C) 1998 by The American Society of Hematology.
引用
收藏
页码:1784 / 1792
页数:9
相关论文
共 65 条
[1]   THE ISOLATION AND CHARACTERIZATION OF 60NM VESICLES (NANOVESICLES) PRODUCED DURING IONOPHORE A23187-INDUCED BUDDING OF HUMAN-ERYTHROCYTES [J].
ALLAN, D ;
THOMAS, P ;
LIMBRICK, AR .
BIOCHEMICAL JOURNAL, 1980, 188 (03) :881-+
[2]   POTOCYTOSIS - SEQUESTRATION AND TRANSPORT OF SMALL MOLECULES BY CAVEOLAE [J].
ANDERSON, RGW ;
KAMEN, BA ;
ROTHBERG, KG ;
LACEY, SW .
SCIENCE, 1992, 255 (5043) :410-411
[3]   BLOOD GROUP-LIKE ACTIVITY RELEASED BY HUMAN MAMMARY-CARCINOMA CELLS IN CULTURE [J].
ANGLIN, JH ;
LERNER, MP ;
NORDQUIST, RE .
NATURE, 1977, 269 (5625) :254-255
[4]   SHEDDING FROM NORMAL AND CANCER-CELL SURFACES [J].
BLACK, PH .
NEW ENGLAND JOURNAL OF MEDICINE, 1980, 303 (24) :1415-1416
[5]  
BRODBECK U, 1981, LAB MANUAL, P85
[6]   PURIFIED GPI-ANCHORED CD4DAF AS A RECEPTOR FOR HIV-MEDIATED GENE-TRANSFER [J].
BRODSKY, RA ;
JANE, SM ;
VANIN, EF ;
MITSUYA, H ;
PETERS, TR ;
SHIMADA, T ;
MEDOF, ME ;
NIENHUIS, AW .
HUMAN GENE THERAPY, 1994, 5 (10) :1231-1239
[7]   SORTING OF GPI-ANCHORED PROTEINS TO GLYCOLIPID-ENRICHED MEMBRANE SUBDOMAINS DURING TRANSPORT TO THE APICAL CELL-SURFACE [J].
BROWN, DA ;
ROSE, JK .
CELL, 1992, 68 (03) :533-544
[8]  
BUTIKOFER P, 1989, BLOOD, V74, P1481
[9]  
Butikofer P, 1997, BIOCHEM J, V326, P415
[10]  
BUTIKOFER P, 1993, J BIOL CHEM, V268, P17794