Expression studies of CLN3 protein (battenin) in fusion with the green fluorescent protein in mammalian cells in vitro.
The gene for Batten disease, the CLN3 gene, encodes a novel, highly hydrophobic, multitransmembrane protein, predicted to consist of 438 amino acid residues. We have expressed a full-length CLN3 protein in fusion with green fluorescent protein in various cell lines to provide its initial biochemical characterization and subcellular localization. By using Western blotting, Percoll density gradient fractionation, and Triton X-114 extraction, we demonstrate that the product of the CLN3 gene, which we call battenin, in mammalian expression system studied is a highly glycosylated protein of lysosomal membrane. In addition our data suggest that CLN3 protein is processed proteolytically in acidic compartments of the cell. Thus, battenin represents the novel constituent of a growing family of lysosomal membrane proteins.
Golabek
A A
AA
Department of Pathological Neurobiology, NYS Institute for Basic Research in Developmental Disabilities, 1050 Forest Hill Road, Staten Island, New York 10314, USA.
Kaczmarski
W
W
Kida
E
E
Kaczmarski
A
A
Michalewski
M P
MP
Wisniewski
K E
KE
D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D002458:Cell Fractionation; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D016213:Cyclins; D011992:Endosomes / Q000378:metabolism; D006031:Glycosylation; D006056:Golgi Apparatus / Q000378:metabolism; D049452:Green Fluorescent Proteins; D066298:In Vitro Techniques; D008164:Luminescent Proteins / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism; D018832:Molecular Chaperones / Q000378:metabolism*; D011233:Precipitin Tests; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D013450:Sulfones / Q000378:metabolism; D013997:Time Factors; D014162:Transfection
C079895:CLN3 protein, S cerevisiae; C105199:CLN3 protein, human; D016213:Cyclins; D008164:Luminescent Proteins; D008562:Membrane Glycoproteins; D018832:Molecular Chaperones; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D013450:Sulfones; D049452:Green Fluorescent Proteins; C002010:4-(2-aminoethyl)benzenesulfonylfluoride