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J. Biol. Chem., Vol. 277, Issue 28, 25592-25600, July 12, 2002

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Molecular Cloning and Characterization of Chick Sialoprotein Associated with Cones and Rods, a Developmentally Regulated Glycoprotein of Interphotoreceptor Matrix^*

Masahiro Zako^§, Masayoshi Iwaki, Masahiko Yoneda^¶, Osamu Miyaishi, Jinsong Zhao, Yasuhiko Suzuki^**, Makoto Takeuchi, Goichiro Miyake, Hiroshi Ikagawa, and Koji Kimata

From the Departments of  Ophthalmology and  Pathology, the  Institute for Molecular Science of Medicine, Aichi Medical University, Nagakute, Aichi 480-1195, the ^¶ Aichi Prefectural College of Nursing and Health Nagoya, Aichi 463-8502, and the ^** Tosei Municipal Hospital, Oiwake, Seto, Aichi 489-0803, Japan

MY-174 is an IgM class monoclonal antibody originally established against chick PG-M/versican. The antibody specifically stains the photoreceptor layer, where we recently reported an absence of PG-M/versican. In this study, we re-characterized the antibody and identified the molecule that reacts to MY-174 at the photoreceptor layer. Immunohistochemistry localized the antigen to the matrix surrounding photoreceptors. A variety of glycosidase digestions showed that the antigen is the 150-kDa glycoprotein that has sialylated N- and O-linked glycoconjugates having a molecular mass of more than 30-kDa. The peptide sequences obtained from purified MY-174 antigen showed we had sequenced a full-length cDNA with an open reading frame of 2787 base pairs, encoding a polypeptide of 928 amino acids, with 56 and 54% identities to human and mouse sialoprotein associated with cones and rods (SPACRs), respectively, and with the structural features observed in SPACRs. The specific sialylated O-glycoconjugates here are involved in the epitope structure for MY-174. SPACR first appeared by embryonic days 15-16, and expression increased with developmental age, paralleling the adhesion between neural retina and retinal pigment epithelium. Thus, we concluded that the MY-174 antigen at the photoreceptor layer, a developmentally regulated glycoprotein, is identical to chick SPACR and may be involved in a novel system mediating adhesion between neural retina and retinal pigment epithelium.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AB070714.

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