We have obtained a monoclonal antibody (33A-3D) that specifically recognize desmoyokin, a 680 kDa desmosomal plaque protein that is well characterized in bovine muzzle epidermis. A cDNA clone (DY6, 3693 bp) was isolated by immunoscreening a mouse keratinocyte expression library with 33A-3D, and it was confirmed that DY6 has a partial coding sequence for desmoyokin. DY6 consists of highly homologous repeats about 128 residues long. Furthermore, the 128-residue repeats exhibit a quasi seven-residue substructure, which we believe will adopt an antiparallel beta-sheet structure. Surprisingly, the amino acid sequence showed a significant homology with AHNAK, a newly identified human gene encoding a 700 kDa protein, which was suggested to be down-regulated in neuroblastoma. From its extensive homology, the similarity in both size and structure, and the identical patterns on Southern blot analysis of genomic DNAs, desmoyokin and AHNAK protein are thought to be identical. Although the desmoyokin/AHNAK protein is detected in a variety of cell types at both protein and mRNA levels, its distribution in keratinocytes (associated closely with cell membrane) is quite different from that in cells other than keratinocytes (distributed diffusely in the cytoplasm). These findings suggest that the desmoyokin/AHNAK protein is a ubiquitous molecule with a unique structure and appears to have different distributions (and probably different functions) among different cells.

Amagai
M.
,
Hashimoto
T.
,
Tajima
S.
,
Inokuchi
Y.
,
Shimizu
N.
,
Saito
M.
,
Miki
K.
,
Nishikawa
T.
(
1990
).
Partial cDNA cloning of the 230-kDa mouse bullous pemphigoid antigen by use of a human monoclonal anti-basement membrane zone antibody.
J. Invest. Dermatol
95
,
252
259
Cohen
S. M.
,
Gorbsky
G.
,
Steinberg
M. S.
(
1983
).
Immunochemical characterization of related families of glycoproteins in desmosomes.
J. Biol. Chem
258
,
2621
2627
Collins
J. E.
,
Legan
P. K.
,
Kenny
T. P.
,
MacGarvie
J.
,
Holton
J. L.
,
Garrod
D. R.
(
1991
).
Cloning and sequence analysis of desmosomal glycoproteins 2 and 3 (Desmocollins): Cadherin-like desmosomal adhesion molecules with heterogeneous cytoplasmic domains.
J. Cell Biol
113
,
381
391
Chou
P. Y.
,
Fasman
G. D.
(
1978
).
Empirical predictions of protein conformation.
Annu. Rev. Biochem
47
,
251
276
Cowin
P.
,
Garrod
D. R.
(
1983
).
Antibodies to epithelial desmosomes show wide tissue and species cross-reactivity.
Nature
302
,
148
150
Cowin
P.
,
Kapprell
H.-P.
,
Franke
W. W.
,
Tamkun
J.
,
Hynes
R. O.
(
1986
).
Plakoglobin: a protein common to different kinds of intercellular adhering junction.
Cell
46
,
1063
1074
Franke
W. W.
,
Goldschmidt
M. D.
,
Zimbelmann
R.
,
Mueller
H. M.
,
Schiller
D. L.
,
Cowin
P.
(
1989
).
Molecular cloning and amino acid sequence of human plakoglobin, the common junctional plaque protein.
Proc. Nat. Acad. Sci. USA
86
,
4027
4031
Franke
W. W.
,
Mueller
H.
,
Mittnacht
S.
,
Kapprell
H.-P.
,
Jorcano
J. J.
) (
1983
).
Significance of two desmosome plaque-associated polypeptides of molecular weights 75000 and 83000.
EMBO J
2
,
2211
2215
Garnier
J.
,
Osguthorpe
D. J.
,
Robson
B.
(
1978
).
Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins.
J. Mol. Biol
120
,
97
120
Green
K. J.
,
Parry
D. A. D.
,
Steinert
P. M.
,
Virata
M. L. A.
,
Wagner
R. M.
,
Angst
B. D.
,
Nilles
L. A.
(
1990
).
Structure of the human desmoplakins. Implication for function in the desmosomal plaque.
J. Biol. Chem
265
,
2603
2612
Gorbsky
G.
,
Steinberg
M. S.
(
1981
).
Isolation of the intercellular glycoproteins of desmosomes.
J. Cell Biol
90
,
243
248
Hashimoto
T.
,
Ogawa
M. M.
,
Konohana
A.
,
Nishikawa
T.
(
1990
).
Detection of pemphigus vulgaris and pemphigus foliaceus antigens by immunoblot analysis using different antigen sources.
J. Invest. Dermatol
94
,
327
331
Hieda
Y.
,
Tsukita
S.
,
Tsukita
S.
(
1989
).
A new high molecular mass protein showing unique localization in desmosomal plaque.
J. Cell Biol
109
,
1511
1518
Kitajima
Y.
,
Inoue
S.
,
Yaoita
H.
(
1989
).
Effect of pemphigus antibody on the regeneration of cell-cell contact in keratinocyte cultures grown in low to normal Ca++ concentration.
J. Invest. Dermatol
89
,
167
171
Koch
P. J.
,
Walsh
M. J.
,
Schmelz
M.
,
Goldschmidt
M. D.
,
Zimbelmann
R.
,
Franke
W. W.
(
1990
).
Identification of desmoglein, a constitutive desmosomal glycoprotein, as a member of the cadherin family of cell adhesion molecules.
Eur.J. Cell Biol
53
,
1
12
Konohana
A.
,
Konohana
I.
,
Roberts
G. P.
,
Marks
R.
(
1987
).
Biochemical changes in desmosomes of bovine muzzle epidermis during differentiation.
J. Invest. Dermatol
89
,
353
357
Laemmli
U. K.
(
1970
).
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature
227
,
680
685
Lipman
D. J.
,
Pearson
W. R.
(
1985
).
Rapid and sensitive protein similarity searches.
Science
227
,
1435
1441
McLachlan
A. D.
,
Stewart
M.
(
1976
).
The 14-fold periodicity in-tropomyosin and interaction with actin.
J. Mol. Biol
103
,
271
298
Mechanic
S.
,
Raynor
K.
,
Hill
J. E.
,
Cowin
P.
(
1991
).
Desmocollins form a distinct subset of the cadherin family of cell adhesion molecule.
Proc. Nat. Acad. Sci. USA
88
,
4476
4480
Merlino
G. T.
,
Ishii
S.
,
Whang-Peng
J.
,
Knutsen
T.
,
Xu
Y.-H.
,
Clark
A. J. L.
,
Stratton
R. H.
,
Wilson
R. K.
,
Ma
D. P.
,
Roe
B. A.
,
Hunts
J. H.
,
Shimizu
N.
,
Pastan
I.
(
1985
).
Structure and localization of genes encoding aberrant and normal growth factor receptor RNAs from A431 human carcinoma cells.
Mol. Cell. Biol
5
,
1722
1734
Moll
R.
,
Cowin
P.
,
Kapprell
H.-P.
,
Franke
W. W.
) (
1986
).
Biology of disease. Desmosomal proteins: New markers for identification and classification of tumors.
Lab. Invest
54
,
4
25
Mueller
H.
,
Franke
W. W.
(
1983
).
Biochemical and immunological characterization of desmoplakins I and II, the major polypeptides of the desmosomal plaque.
J. Mol. Biol
163
,
647
671
Nilles
L. A.
,
Parry
D. A. D.
,
Powers
E. E.
,
Angst
B. D.
,
Wagner
R. M.
,
Green
K. J.
(
1991
).
Structural analysis and expression of human desmoglein: A cadherin-like component of the desmosome.
J. Cell Sci
99
,
809
821
O'Keefe
E. J.
,
Erickson
H. P.
,
Bennett
V.
(
1989
).
Desmoplakin I and desmoplakin II. Function and characterization.
J. Biol. Chem
264
,
8310
8318
Parker
A. E.
,
Wheeler
G. N.
,
Arnemann
J.
,
Pidsley
S. C.
,
Ataliotis
P.
,
Thomas
C. L.
,
Rees
D. A.
,
Magee
A. I.
,
Buxton
R. S.
(
1991
).
Desmosomal glycoproteins II and III. Cadherin-like junctional molecules generated by alternative splicing.
J. Biol. Chem
266
,
10438
10445
Parrish
E. P.
,
Marston
J. E.
,
Mattey
D. L.
,
Measures
H. R.
,
Venning
R.
,
Garrod
D. R.
(
1990
).
Size heterogeneity, phosphorylation and transmembrane organization of desmosomal glycoproteins 2 and 3 (desmocollins) in MDCK cells.
J. Cell Sci
96
,
239
248
Parrish
E. P.
,
Steart
P. V.
,
Garrod
D. R.
,
Weller
R. O.
(
1987
).
Antidesmsomal mAb in the diagnosis of intracranial tumors.
J. Pathol
153
,
265
273
Parry
D. A. D.
(
1975
).
Analysis of the primary structure of-tropomyosin from rabbit skeletal muscle.
J. Mol. Biol
98
,
519
535
Richardson
J. S.
(
1981
).
The anatomy and taxonomy of protein structure.
Advan. Prot. Chem
34
,
167
339
Saito
H.
,
Morizane
T.
,
Watanabe
T.
,
Kagawa
T.
,
Iwabuchi
N.
,
Kumagai
N.
,
Inagaki
Y.
,
Tsuchimoto
K.
,
Tsuchiya
M.
(
1989
).
Establishment of a human cell line (HCC-T) from a patient with hepatoma bearing no evidence of hepatitis B or A virus infection.
Cancer
64
,
1054
1060
Sanger
F.
,
Nicklen
S.
,
Coulson
A. R.
(
1977
).
DNA sequencing with chain-terminating inhibitors.
Proc. Nat. Acad. Sci. USA
74
,
5463
5467
Schmelz
M.
,
Duden
R.
,
Cowin
P.
,
Franke
W. W.
(
1986
).
A constitutive transmembrane glycoprotein of Mr 165000 (desmoglein) in epidermal and non-epidermal desmosomes. I. Biochemical identification of the polypeptide.
Eur. J. Cell Biol
42
,
177
183
Schwarz
M. A.
,
Owaribe
K.
,
Kartenbeck
J.
,
Franke
W. W.
(
1990
).
Desmosomes and hemidesmosomes: constitutive molecular components.
Annu. Rev. Cell Biol
6
,
461
491
Shtivelman
E.
,
Cohen
F. E.
,
Bishop
J. M.
(
1992
).
A human gene (AHNAK) encoding an unusually large protein with a 1.2-m polyionic rod structure.
Proc. Nat. Acad. Sci. USA
89
,
5472
5476
Skerrow
C. J.
,
Matoltsy
A. G.
(
1974
).
Isolation of epidermal desmosomes.
J. Cell Biol
63
,
515
523
Smith
D. B.
,
Johnson
K. S.
(
1988
).
Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase.
Gene
67
,
31
40
Tsukamoto
T.
(
1989
).
Establishment and characterization of a cell line (KU-8) from squamous cell carcinoma of the penis.
Keio J. Med
38
,
277
293
Tsukita
S.
,
Tsukita
S.
(
1985
).
Desmocalmin: A calmodulin-binding high molecular weight protein isolated from desmosomes.
J. Cell Biol
98
,
1565
1571
Virata
M. L. A.
,
Wagner
R. M.
,
Parry
D. A. D.
,
Green
K. J.
(
1992
).
Molecular structure of the human desmoplakins I and II amino terminus.
Proc. Nat. Acad. Sci. USA
89
,
544
548
Watt
F. M.
,
Mattey
D. L.
,
Garrod
D. R.
(
1984
).
Calcium induced reorganization of desmosomal components in cultured human keratinocytes.
J. Cell Biol
99
,
2211
2215
Wheeler
G. N.
,
Parker
A. E.
,
Thomas
C. L.
,
Ataliotes
P.
,
Poynter
D.
,
Arnemann
J.
,
Rutman
A. J.
,
Pidsley
S. C.
,
Watt
F. M.
,
Rees
D. A.
,
Buxton
R. S.
,
Magee
A. I.
(
1990
).
Desmosomal glycoprotein DGI, a component of intercellular desmosome junctions, is related to the cadherin family of cell adhesion molecules.
Proc. Nat. Acad. Sci. USA
88
,
4796
4880
Wiche
G.
,
Becker
B.
,
Luber
K.
,
Weitzer
G.
,
Castanon
M. J.
,
Hauptmann
R.
,
Stratowa
C.
,
Stewart
M.
(
1991
).
Cloning and sequencing of Rat Plectin indicates a 466-kDa polypeptide chain with a three-domain structure based on a central alpha-helical coiled coil.
J. Cell Biol
114
,
83
99
Wiche
G. R.
,
Krepler
R.
,
Artlieb
U.
,
Pytela
R.
,
Denk
H.
(
1983
).
Occurrence and immunolocalization of plectin in tissues.
J. Cell Biol
97
,
887
901
Yanisch-Perron
C.
,
Vieira
J.
,
Messing
J.
(
1985
).
Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors.
Gene
33
,
103
119
Yuspa
S. H.
,
Hawley-Nelson
P.
,
Koehler
B.
,
Stanley
J. R.
(
1980
).
Survey of transformation markers in differentiating epidermal cell lines in culture.
Cancer Res
40
,
4694
4703
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