Molecular characterization of Golgin-245, a novel Golgi complex protein containing a granin signature.
The serum from a Sjögren's syndrome patient with anti-Golgi antibodies was used as a probe to isolate a 4.6-kilobase pair cDNA insert from a HeLa cDNA library. Expression of the cDNA in Escherichia coli and the in vitro translation products of the cDNA yielded a recombinant protein that migrated in SDS-polyacrylamide gel electrophoresis at 180 kDa. This protein was immuno-precipitated by the human anti-Golgi serum and by immune rabbit serum but not by normal human serum or preimmune rabbit serum. Western blot analysis showed that the prototype human and immune rabbit sera recognized a 245-kDa protein, suggesting that the isolated clone contained a partial cDNA. The 5'-upstream sequence obtained by the rapid amplification of cDNA ends methodology using human placental cDNA and the combined HeLa cDNA contained 6965 base pairs and combined HeLa cDNA contained 6965 base pairs and encoded a protein of 245 kDa and, like other Golgi autoantigens described earlier, is highly rich in coiled-coils. The deduced amino acid sequence included the decapeptide ESLALEELEL, which was identified as one of two signature sequences previously reported in a family of peptide hormones and neuropeptides known as "granins". This is the first report of a Golgi complex autoantigen that bears structural similarities to the granin family of proteins.
Department of Medical Biochemistry, University of Calgary, Alberta, Canada.
M J
MJ
Fritzler
C C
CC
Lung
J C
JC
Hamel
K J
KJ
Griffith
E K
EK
Chan
D000595:Amino Acid Sequence; D000818:Animals; D001324:Autoantigens / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D002864:Chromogranins / Q000737:chemistry*; D003001:Cloning, Molecular; D018076:DNA, Complementary; D006056:Golgi Apparatus / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D011817:Rabbits; D012859:Sjogren's Syndrome / Q000097:blood
D001324:Autoantigens; D002864:Chromogranins; D018076:DNA, Complementary; C098772:GOLGA4 protein, human; D008565:Membrane Proteins