CORRECTLY SORTED MOLECULES OF A GPI-ANCHORED PROTEIN ARE CLUSTERED AND IMMOBILE WHEN THEY ARRIVE AT THE APICAL SURFACE OF MDCK CELLS

被引:94
作者
HANNAN, LA [1 ]
LISANTI, MP [1 ]
RODRIGUEZBOULAN, E [1 ]
EDIDIN, M [1 ]
机构
[1] CORNELL UNIV, SCH MED, DEPT CELL BIOL & ANAT, NEW YORK, NY 10021 USA
关键词
D O I
10.1083/jcb.120.2.353
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Glycosyl-phosphatidylinositol (GPI)-anchored proteins are sorted to the apical surface of many epithelial cell types. To better understand the mechanism for apical segregation of these proteins, we analyzed the lateral mobility and molecular associations of a model GPI-anchored protein, herpes simplex virus gD1 fused to human decay accelerating factor (gD1-DAF) (Lisanti, M. P., I. W. Caras, M. A. Davitz, and E. Rodriguez-Boulan. 1989. J. Cell Biol. 109:2145-2156) shortly after arrival and after long-term residence at the surface of confluent, polarized MDCK cells. FRAP measurements of lateral diffusion showed that the mobile fraction of newly arrived gD1-DAF molecules was much less than the mobile fraction of long-term resident molecules (40 vs. 80-90%). Fluorescence resonance energy transfer measurements showed that the newly arrived molecules were clustered, while resident molecules were not. Newly delivered gD1-DAF molecules were clustered but not immobilized in mutant, Concanavalin A-resistant MDCK cells that failed to sort gD1-DAF. Our results indicate that GPI-anchored proteins in MDCK cells are clustered before delivery to the surface. However, clustering alone does not target molecules for apical delivery. The immobilization observed when gD1-DAF is correctly sorted suggests that the clusters must associate some component of the cell's cytoplasm.
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页码:353 / 358
页数:6
相关论文
共 53 条
[1]   VESICLES AND CISTERNAE IN THE TRANS GOLGI-APPARATUS OF HUMAN-FIBROBLASTS ARE ACIDIC COMPARTMENTS [J].
ANDERSON, RGW ;
PATHAK, RK .
CELL, 1985, 40 (03) :635-643
[2]   MOBILITY MEASUREMENT BY ANALYSIS OF FLUORESCENCE PHOTOBLEACHING RECOVERY KINETICS [J].
AXELROD, D ;
KOPPEL, DE ;
SCHLESSINGER, J ;
ELSON, E ;
WEBB, WW .
BIOPHYSICAL JOURNAL, 1976, 16 (09) :1055-1069
[3]   LATERAL MOBILITY OF CLASS-I HISTOCOMPATIBILITY ANTIGENS IN B-LYMPHOBLASTOID CELL-MEMBRANES - MODULATION BY CROSS-LINKING AND EFFECT OF CELL-DENSITY [J].
BIERER, BE ;
HERRMANN, SH ;
BROWN, CS ;
BURAKOFF, SJ ;
GOLAN, DE .
JOURNAL OF CELL BIOLOGY, 1987, 105 (03) :1147-1152
[4]   A SINGLE AMINO-ACID CHANGE IN THE CYTOPLASMIC DOMAIN ALTERS THE POLARIZED DELIVERY OF INFLUENZA-VIRUS HEMAGGLUTININ [J].
BREWER, CB ;
ROTH, MG .
JOURNAL OF CELL BIOLOGY, 1991, 114 (03) :413-421
[5]   MECHANISM OF MEMBRANE ANCHORING AFFECTS POLARIZED EXPRESSION OF 2 PROTEINS IN MDCK CELLS [J].
BROWN, DA ;
CRISE, B ;
ROSE, JK .
SCIENCE, 1989, 245 (4925) :1499-1501
[6]   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
[7]   DEPENDENCE ON PH OF POLARIZED SORTING OF SECRETED PROTEINS [J].
CAPLAN, MJ ;
STOW, JL ;
NEWMAN, AP ;
MADRI, J ;
ANDERSON, HC ;
FARQUHAR, MG ;
PALADE, GE ;
JAMIESON, JD .
NATURE, 1987, 329 (6140) :632-635
[8]   SIGNAL FOR ATTACHMENT OF A PHOSPHOLIPID MEMBRANE ANCHOR IN DECAY ACCELERATING FACTOR [J].
CARAS, IW ;
WEDDELL, GN ;
DAVITZ, MA ;
NUSSENZWEIG, V ;
MARTIN, DW .
SCIENCE, 1987, 238 (4831) :1280-1283
[9]   SIGNAL PEPTIDE FOR PROTEIN SECRETION DIRECTING GLYCOPHOSPHOLIPID MEMBRANE ANCHOR ATTACHMENT [J].
CARAS, IW ;
WEDDELL, GN .
SCIENCE, 1989, 243 (4895) :1196-1198
[10]  
CASANOVA JE, 1991, J BIOL CHEM, V266, P24428