Human amyloid precursor-like protein 1--cDNA cloning, ectopic expression in COS-7 cells and identification of soluble forms in the cerebrospinal fluid.
Amyloid precursor-like protein 1 (APLP1) represents an integral membrane type 1 protein of unknown function which was originally cloned from a mouse cDNA library on the basis of sequence similarity with the Alzheimer's amyloid precursor protein (APP). Here we report on the molecular cloning and expression of the human APLP1 (hAPLP1). hAPLP1 consists of 650 amino acids, displays 89% identity on the amino acid level to its mouse homologue and has a calculated molecular mass of 72 kDa. hAPLP1 synthesized in a cell-free system displays an apparent molecular mass of approximately 80 kDa in SDS-containing gels and becomes N-glycosylated when the in vitro translation is performed in the presence of microsomes. The hAPLP1 cDNA was also expressed ectopically in COS-7 cells and the protein expression was analyzed by immunoprecipitation and western blotting. We have demonstrated that hAPLP1 represents a novel glycoprotein which carries both N- and O-linked glycans. Moreover, hAPLP1 undergoes limited proteolysis which results in the secretion of the carboxy-terminal truncated molecule into the cells conditioned medium. Examination of cells transfected with hAPLP1 cDNA by confocal laser microscopy reveals an intense perinuclear and Golgi staining, a pattern resembling the subcellular distribution of APP. Using a novel hAPLP1-specific antiserum, we identified soluble hAPLP1 in the human cerebrospinal fluid, which suggests that secretion of hAPLP1 from brain cells also takes place in vivo.
Paliga
K
K
Center for Molecular Biology Heidelberg, Germany. krzych@sun0.urz.uni-heidelberg.de
Peraus
G
G
Kreger
S
S
Dürrwang
U
U
Hesse
L
L
Multhaup
G
G
Masters
C L
CL
Beyreuther
K
K
Weidemann
A
A
D000595:Amino Acid Sequence; D016564:Amyloid beta-Protein Precursor / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000134:cerebrospinal fluid / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D003001:Cloning, Molecular; D006801:Humans; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D011817:Rabbits; D014158:Transcription, Genetic
C489697:APLP1 protein, human; D016564:Amyloid beta-Protein Precursor