COMPARATIVE BIOCHEMICAL-ANALYSIS OF PURIFIED S-ANTIGEN FROM HUMAN, BOVINE, PORCINE AND RAT RETINA

被引:17
作者
BANGA, JP [1 ]
KASP, E [1 ]
SULEYMAN, S [1 ]
BROWN, E [1 ]
ELLIS, BA [1 ]
SANDERS, MD [1 ]
DUMONDE, DC [1 ]
机构
[1] UNITED MED & DENT SCH GUYS & ST THOMAS HOSP, DEPT IMMUNOL & MED OPHTHALMOL, ST THOMAS CAMPUS, LONDON SE1 7EH, ENGLAND
关键词
D O I
10.1016/S0014-4835(87)80004-0
中图分类号
R77 [眼科学];
学科分类号
100212 ;
摘要
Highly purified S-antigen from human, bovine, porcine and rat retina was used to ascertain the molecular weights, isoelectric points, amino-acid composition and presence of carbohydrate moieties. All S-antigen preparations comigrate to 50,000 MW on SDS-polyacrylamide gels and demonstrate microheterogeneity upon isoelectric focusing. Human S-antigen exhibits two bands at ph 5.9 and 5.6, whilst with bovine S-antigen a broad band at pH 5.9 was observed. Porcine and rat S-antigen gave four bands, focusing between pH 5.0-5.6. Two-dimensional gel analysis revealed that all the four focused porcine polypeptides migrate to 50,000 MW. Western-blot analysis following isoelectric focusing of human and porcine S-antigen showed that all polypeptide bands react specifically with polyclonal and monoclonal antibodies to S-antigen. The reasons for the apparent heterogeneity of S-antigen by isolectric focusing are discussed. All S-antigen preparations were weakly positive for periodic acid Schiff staining, and specifically bound radiolabelled Lens culinaris lectin on Western-blot analysis. These results confirm the glycoprotein nature of S-antigen which may explain the finding of multiple bands found on isolectric focusing. Amino-acid analysis reveals the high percentage of non-polar amino acids and may explain the apparent hydrophobic behaviour of the isolated protein.
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页码:199 / 207
页数:9
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