CENP-C, an autoantigen in scleroderma, is a component of the human inner kinetochore plate.
We have isolated and characterized a set of overlapping cDNA clones that encode the human centromere autoantigen centromere protein C (CENP-C). The identity of these clones has been established using several criteria. First, they were shown to encode a polypeptide that migrates at the expected position for CENP-C on SDS-polyacrylamide gel electrophoresis. Second, we have demonstrated that this polypeptide shares at least two epitopes with human CENP-C. Polyclonal antibodies were raised to fusion proteins encoded by nonoverlapping regions of the cDNA clones. These antibodies were shown to recognize a protein at a position appropriate for CENP-C on immunoblots of human chromosomal proteins. In addition, we used indirect immunofluorescence to demonstrate that these antibodies recognize centromeres of HeLa chromosomes in the expected pattern for CENP-C. Localization of CENP-C by immunoelectron microscopy reveals that this protein is a component of the inner kinetochore plate.
Saitoh
H
H
Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Tomkiel
J
J
Cooke
C A
CA
Ratrie
H
H
Maurer
M
M
Rothfield
N F
NF
Earnshaw
W C
WC
D000595:Amino Acid Sequence; D001324:Autoantigens / Q000737:chemistry*; D001483:Base Sequence; D002503:Centromere / Q000737:chemistry*; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis*; D002875:Chromosomes / Q000737:chemistry; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D011928:Raynaud Disease / Q000235:genetics* / Q000276:immunology
D001324:Autoantigens; D002868:Chromosomal Proteins, Non-Histone; C060905:centromere protein C; D004247:DNA