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J. Biochem, 1988, Vol. 104, No. 4 658-666
© 1988 Japanese Biochemical Society

research-article

The Localization of Sulfated Glycoconjugates Synthesized by the Corneal Epithelium of Chick Embryos1

Hideto Yonekura**,2, Kayoko Oguri*, Kazuhiro Itoh*, Yasuo Nakanishi** and Minoru Okayama*,3

*Clinical Research Institute, National Nagoya Hospital Naka-ku, Nagoya, Aichi 460
**Department of Chemistry, Faculty of Science, Nagoya University Chikusa-ku, Nagoya, Aichi 464

3To whom correspondence should be addressed

The localization of sulfated glycoconjugates in the corneal epithelium of 19.d-old chick embryo was investigated biochemically using epithelia labeled in vitro with [35 sulfate, which exhibited autoradiographically a similar distribution of silver grains to that labeled in ovo. The radiolabeled tissues were dissociated into single cells by incubation in 0.25% trypsin containing 0.0%EDTA at 37 for 40 min. The proteoglycans and sulfated glycoproteins which were associated with the cells and those released into the dissociation medium were separated by DEAE-Sepharose CL-6B and analyzed on Sepharose CL-6B and SDS-PAGE. About 86% of the proteoglycans was released into the dissociation medium and more than 50% of the cell-associated ones was affected by trypsin. This indicates that the proteoglycans are mostly localized in an extracellular compartment. On the other hand, the extent of release of sulfated glycoproteins into the medium on dissociation of tissues was distinctly different depending upon their molecular weight (Mr): almost all of the sulfated glycoproteins of the family with Mr 48,000–70,000(32% of the total sulfated glycoproteins) were recovered as intact molecules with the cells, whereas approximately 50% of those with Mr 70,000–150,000 (36%) and about 70% of those with Mr over 150,000 (28%) were released into the dissociation medium. These results indicate that the family with Mr 48,000–70,000 is localized intracellularly and that with Mr 70,000–150,000 in a compartment poorly affected by trypsin; in contrast to those, that with Mr more than 150,000 is localized in an extracellular compartment like the proteoglycans.

1This study was supported in part by a Grant-in-Aid for Cancer Research (60-4) from the Ministry of Health and Welfare of Japan, and by grants from the Foundation for Promotion of Cancer Research and The Fujisawa Foundation.

2Present address: Cell Engineering Creative Products Research Laboratories Research Institute, Kissei Pharmaceutical Co., Ltd, Matsumoto, Nagano 399.


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