Further characterization of human mucin-like glycoprotein associated with photoreceptor cells by its introduction into Y79 retinoblastoma cells.
OBJECTIVE
To understand the function of a mucin- like glycoprotein associated with photoreceptor cells (MLGAPC), MLGAPC-transfected Y79 retinoblastoma cells were analyzed biochemically and morphologically.


METHODS
The cDNA for the core protein of human MLGAPC was isolated, cloned into a mammalian expression vector, pCAGGSneo, and then transfected into Y79 retinoblastoma cells. The cells and the medium were analyzed by means of Western blotting, and the morphology of the transfectants and parental cells was compared.


RESULTS
Western blot analysis of the culture medium revealed that the transfectants secreted MLGAPC into the medium. Lectin blot analysis of MLGAPC in the medium showed that it had a binding site for Maackia amurensis lectin II. No morphological difference was detected between the transfectants and parental cells.


CONCLUSIONS
As expected from the deduced amino acid sequence of MLGAPC, it was secreted into the medium. The secreted MLGAPC was found to carry sialoglycans (rod type). The expression of MLGAPC had no effect on the adhesion or morphology of the cultured cells, which suggests that its interaction with other components may be required for these effects.
Yamashita
Takehiro
T
Department of Ophthalmology, Kagoshima University, Faculty of Medicine, Kagoshima, Japan.
Uehara
Fumiyuki
F
Ozawa
Masayuki
M
Ohba
Norio
N
D001665:Binding Sites; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification; D016326:Extracellular Matrix Proteins; D005136:Eye Proteins / Q000502:physiology*; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D006023:Glycoproteins / Q000502:physiology*; D006801:Humans; D015151:Immunoblotting; D037102:Lectins / Q000378:metabolism; D009077:Mucins / Q000502:physiology*; D020419:Photoreceptor Cells, Vertebrate / Q000378:metabolism*; D011509:Proteoglycans; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D019572:Retinal Neoplasms / Q000378:metabolism* / Q000473:pathology; D012175:Retinoblastoma / Q000378:metabolism* / Q000473:pathology; D014162:Transfection; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D016326:Extracellular Matrix Proteins; D005136:Eye Proteins; D006023:Glycoproteins; C486428:IMPG1 protein, human; D037102:Lectins; D009077:Mucins; D011509:Proteoglycans; D011993:Recombinant Fusion Proteins