VIP36 was isolated from MDCK cells as a component of glycolipid-enriched detergent-insoluble complexes. The protein is localized to the Golgi apparatus and the cell surface, and belongs to a new family of legume lectin homologues in the animal secretory pathway that might be involved in the trafficking of glycoproteins, glycolipids or both. Here we show that VIP36 is N-glycosylated and expressed in organs abundant in epithelial cells as well as in non-epithelial organs. Our studies demonstrate that the recombinant exoplasmic/luminal domain of VIP36 binds Ca2+ and that the protein decorates internal membrane structures of MDCK cells in vitro that are distinct from the Golgi apparatus. This binding requires Ca2+ and can be specifically inhibited by N-acetyl-D-galactosamine. The recombinant protein was used for affinity chromatography. Glycopeptides obtained from [3H]galactose-labelled cells bind to VIP36 and can be eluted with N-acetyl-D-galactosamine. Our data imply that VIP36 functions as a lectin in post-Golgi trafficking.

REFERENCES

Arar
C.
,
Carpentier
V.
,
Le Caer
J.-P.
,
Monsigny
M.
,
Legrand
A.
,
Roche
A.-C.
(
1995
).
ERGIC-53, a membrane protein of the endoplasmic reticulum-golgi intermediate compartment, is identical to MR60, an intracellular mannose-specific lectin of myelomonocytic cells.
J. Biol. Chem
270
,
3551
3553
Bennett
M.
,
Wandinger-Ness
A.
,
Simons
K.
(
1988
).
Release of putative exocytic transport vesicles from perforated MDCK cells.
EMBO J
7
,
4075
4085
Carpentier
V.
,
Vassard
C.
,
Plessis
C.
,
Motta
G.
,
Monsigny
M.
,
Roche
A. C.
(
1994
).
Characterization and cellular localization by monoclonal antibodies of the 60 kDa mannose specific lectin of human promyelocytic cells, HL60.
Glycoconjugate J
11
,
333
338
Chavrier
P.
,
Simons
K.
,
Zerial
M.
(
1992
).
The high complexity of the rab and rho GTP-binding protein subfamilies revealed by a PCR cloning approach.
Gene
112
,
261
264
Evan
G. I.
,
Lewis
G. K.
,
Ramsay
G.
,
Bishop
J. M.
(
1985
).
Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product.
Mol. Cell. Biol
5
,
3610
3616
Fiedler
K.
,
Kobayashi
T.
,
Kurzchalia
T. V.
,
Simons
K.
(
1993
).
Glycosphingolipid-enriched detergent-insoluble complexes in protein sorting in epithelial cells.
Biochemistry
32
,
6365
6373
Fiedler
K.
,
Parton
R. G.
,
Kellner
R.
,
Etzold
T.
,
Simons
K.
(
1994
).
VIP36, a novel component of glycolipid rafts and exocytic carrier vesicles in epithelial cells.
EMBO J
13
,
1729
1740
Fiedler
K.
,
Simons
K.
(
1994
).
A putative novel class of animal lectins in the secretory pathway homologous to leguminous lectins.
Cell
77
,
625
626
Fiedler
K.
,
Lafont
F.
,
Parton
R. G.
,
Simons
K.
(
1995
).
Annexin XIIIb: a novel epithelial specific annexin is implicated in vesicular traffic to the apical plasma membrane.
J. Cell Biol
128
,
1043
1053
Fiedler
K.
,
Simons
K.
(
1995
).
The role of N-glycans in the secretory pathway.
Cell
81
,
309
312
Kreis
T. E.
(
1986
).
Microinjected antibodies against the cytoplasmic domain of vesicular stomatitis virus glycoprotein, block its transport to the cell surface.
EMBO J
5
,
931
941
Lis
H.
,
Sharon
N.
(
1986
).
Lectins as molecules and as tools.
Annu. Rev. Biochem
55
,
35
67
Lutcke
A.
,
Jansson
S.
,
Parton
R. G.
,
Chavrier
P.
,
Valencia
A.
,
Huber
L. A.
,
Lehtonen
E.
,
Zerial
M.
(
1993
).
Rab17, a novel small GTPase, is specific for epithelial cells and is induced during cell polarization.
J. Cell Biol
121
,
553
564
Matter
K.
,
Mellman
I.
(
1994
).
Mechanisms of cell polarity: Sorting and transport in epithelial cells.
Curr. Opin. Cell Biol
6
,
545
554
Merkle
R. K.
,
Cummings
R. D.
(
1987
).
Lectin affinity chromatography of glycopeptides.
Meth. Enzymol
138
,
232
259
Pimpaneau
V.
,
Midoux
P.
,
Monsigny
M.
,
Roche
A. C.
(
1991
).
Characterization and isolation of an intracellular D-mannose-specific receptor from human promyelocytic HL60 cells.
Carbohydrate Res
213
,
95
108
Rodriguez-Boulan
E.
,
Powell
S. K.
(
1992
).
Polarity of epithelial and neuonal cells.
Annu. Rev. Cell Biol
8
,
395
427
Schindler
R.
,
Itin
C.
,
Zerial
M.
,
Lottspeich
F.
,
Hauri
H.-P.
(
1993
).
ERGIC-53, a membrane protein of the ER-Golgi intermediate compartment, carries an ER retention motif.
Eur. J. Cell Biol
61
,
1
9
Schweizer
A.
,
Fransen
J. A. M.
,
Bachi
T.
,
Ginsel
L.
,
Hauri
H. P.
(
1988
).
Identification, by a monoclonal antibody, of a 53-kD protein associated with a tubular vesicular compartment at the cis-side of the Golgi apparatus.
J. Cell Biol
107
,
1643
1653
Sharon
N.
,
Lis
H.
(
1990
).
Legume lectins- a large family of homologous proteins.
FASEB J
4
,
3198
3208
Simons
K.
,
Wandinger-Ness
A.
(
1990
).
Polarized sorting in epithelia.
Cell
62
,
207
210
Wandinger-Ness
A.
,
Bennet
M. K.
,
Antony
C.
,
Simons
K.
(
1990
).
Distinct transport vesicles mediate the delivery of plasma membrane proteins to the apical and basolateral domains of MDCK cells.
J. Cell Biol
111
,
987
1000
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