Photoaffinity labeling of the lysosomal neuraminidase from bovine testis

FEBS Letters

Rose, Ursula¹  Brossmer, Reinhard¹  van der Horst, Gijsbertus T.J.²  Verheijen, Frans W.² 

[1] Department of Biochemistry II, University of Heidelberg, Heidelberg, Germany;Department of Cell Biology and Genetics, Erasmus University, Rotterdam, The Netherlands

关键词: Neuraminidase;    Sialidase;    Lysosomal;    Photoaifinity labeling;    ASA-NeuAc2en;    5-N-acetyl-9-(4-azidosalicoylamido)-2-deoxy-2;    3-didehydroneuraminic acid;    IASA-NeuAc2en;    5-N-acetyl-9-(4-iodoazidosalicoylamido)-2-deoxy-2;    3-didehydroneuraminie acid;    NeuAc2en;    2-deoxy-2;    3-didehydro-5-N-acetylneuraminic acid;    MU-NeuAc;    2-α'-(4-methylumbelliferyl)-5-N-acetylneuraminic acid;    SDS;    sodium dodecyl sulfate;   

DOI  :  10.1016/0014-5793(90)80805-S

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

ASA-NeuAc2en, a photoreactive arylazide derivative of sialic acid, is shown to be a powerful competitive inhibitor of lysosomal neuraminidase from bovine testis (K _i ≈ 21 μM). Photoaffinity labeling and partial purification of preparations containing this lysosomal neuraminidase activity result in specifically and non-specifically labeled polypeptides. Only labeling in a 55 kDa polypeptide is found to be specific, since it could be prevented by the competitive neuraminidase inhibitor NeuAc2en. We conclude that the 55 kDa polypeptide in the bovine testis β-galactosidase/neuraminidase/protective protein complex contains the catalytic site of neuraminidase.

【 授权许可】

Unknown