Molecular properties and chromosomal location of cadherin-8.
Cloning of rat cadherin-8 cDNA demonstrated two types of cDNAs. The overall structure of the protein defined by one type of the cDNA is essentially the same as that of classic cadherins, whereas the protein defined by the other type of cDNA ends near the N-terminus of the fifth repeat of the extracellular domain (EC5) and contains a short unique sequence at the C-terminus. The same truncated type of cDNA was also obtained from a human cDNA library. In Northern blot analysis of rat brain mRNA, a probe for EC5 detected multiple bands of about 3.5-4.3 knt, whereas a probe for the alternative form hybridized with a band of about 3.5 knt. Western blot experiments showed that an antibody against the extracellular domain of rat cadherin-8 stained a band of about 95 kDa and a faint band of about 130 kDa in rat brain extract. These results suggest that cadherin-8 is expressed in two forms, a complete form and a truncated form without a transmembrane domain or cytoplasmic domain, in brain. The complete form of cadherin-8 expressed in L cells was about 130 kDa in molecular mass and was located at the cell periphery, mainly at the cell-cell contact sites. However, we failed to express the truncated form in L cells. The transfectants of the complete form showed weak cell adhesion activity. The complete form of cadherin-8 was sensitive to trypsin digestion, and Ca2+ did not protect cadherin-8 from digestion, in contrast to the classic cadherins. The complete form of cadherin-8 coprecipitated with beta-catenin, but did not immunoprecipitate well with alpha-catenin or gamma-catenin. Cadherin-8, as well as cadherin-11, was mapped to a specific region of chromosome 8 that also includes cadherins-1, -3, and -5.
Kido
M
M
Doheny Eye Institute, University of Southern California School of Medicine, Los Angeles 90033, USA.
Obata
S
S
Tanihara
H
H
Rochelle
J M
JM
Seldin
M F
MF
Taketani
S
S
Suzuki
S T
ST
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism; D015820:Cadherins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D003433:Crosses, Genetic; D015870:Gene Expression; D006801:Humans; D007739:L Cells; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D051381:Rats; D017422:Sequence Analysis, DNA; D014162:Transfection
C112316:CDH8 protein, human; D015820:Cadherins; C504541:Cdh8 protein, mouse; C504540:Cdh8 protein, rat