Human amyloid precursor-like protein 1 - cDNA cloning, ectopic expression in COS-7 cells and identification of soluble forms in the cerebrospinal fluid

被引:51
作者
Paliga, K [1 ]
Peraus, G [1 ]
Kreger, S [1 ]
Durrwang, U [1 ]
Hesse, L [1 ]
Multhaup, G [1 ]
Masters, CL [1 ]
Beyreuther, K [1 ]
Weidemann, A [1 ]
机构
[1] UNIV MELBOURNE, DEPT PATHOL, MELBOURNE, VIC, AUSTRALIA
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1997年 / 250卷 / 02期
关键词
amyloid precursor-like protein 1; Alzheimer; glycosylation; secretion; cerebrospinal fluid;
D O I
10.1111/j.1432-1033.1997.0354a.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 approximate to 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.
引用
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页码:354 / 363
页数:10
相关论文
共 47 条
[1]  
ADAMS MD, 1995, NATURE, V377, P3
[2]   Increased gene expression of beta-amyloid precursor protein and its homologues APLP1 and APLP2 in human neuroblastoma cells in response to retinoic acid [J].
Beckman, M ;
Iverfeldt, K .
NEUROSCIENCE LETTERS, 1997, 221 (2-3) :73-76
[3]   cDNA cloning and chromosome mapping of the human Fe65 gene: Interaction of the conserved cytoplasmic domains of the human beta-amyloid precursor protein and its homologues with the mouse Fe65 protein [J].
Bressler, SL ;
Gray, MD ;
Sopher, BL ;
Hu, QB ;
Hearn, MG ;
Pham, DG ;
Dinulos, MB ;
Fukuchi, KI ;
Sisodia, SS ;
Miller, MA ;
Disteche, CM ;
Martin, GM .
HUMAN MOLECULAR GENETICS, 1996, 5 (10) :1589-1598
[4]  
Crain BJ, 1996, AM J PATHOL, V149, P1087
[5]   MEMBRANE-PROTEINS WITH SOLUBLE COUNTERPARTS - ROLE OF PROTEOLYSIS IN THE RELEASE OF TRANSMEMBRANE PROTEINS [J].
EHLERS, MRW ;
RIORDAN, JF .
BIOCHEMISTRY, 1991, 30 (42) :10065-10074
[6]   A TECHNIQUE FOR RADIOLABELING DNA RESTRICTION ENDONUCLEASE FRAGMENTS TO HIGH SPECIFIC ACTIVITY [J].
FEINBERG, AP ;
VOGELSTEIN, B .
ANALYTICAL BIOCHEMISTRY, 1983, 132 (01) :6-13
[7]   The regions of the Fe65 protein homologous to the phosphotyrosine interaction phosphotyrosine binding domain of Shc bind the intracellular domain of the Alzheimer's amyloid precursor protein [J].
Fiore, F ;
Zambrano, N ;
Minopoli, G ;
Donini, V ;
Duilio, A ;
Russo, T .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (52) :30853-30856
[8]   Membrane protein secretases [J].
Hooper, NM ;
Karran, EH ;
Turner, AJ .
BIOCHEMICAL JOURNAL, 1997, 321 :265-279
[9]   LECTIN AFFINITY-CHROMATOGRAPHY OF PROTEINS BEARING O-LINKED OLIGOSACCHARIDES - APPLICATION OF JACALIN-AGAROSE [J].
HORTIN, GL ;
TRIMPE, BL .
ANALYTICAL BIOCHEMISTRY, 1990, 188 (02) :271-277
[10]   THE PRECURSOR OF ALZHEIMERS-DISEASE AMYLOID-A4 PROTEIN RESEMBLES A CELL-SURFACE RECEPTOR [J].
KANG, J ;
LEMAIRE, HG ;
UNTERBECK, A ;
SALBAUM, JM ;
MASTERS, CL ;
GRZESCHIK, KH ;
MULTHAUP, G ;
BEYREUTHER, K ;
MULLERHILL, B .
NATURE, 1987, 325 (6106) :733-736