Post‐translational processing and Thr‐206 are required for glycosylasparaginase activity

FEBS Letters

Vettese, Michelle B.¹  Klein, Melissa¹  Fisher, Krishna J.¹  Park, Hyejeong¹  Aronson, Nathan N.¹ 

[1] Department of Molecular and Cell Biology, Althouse Lab., The Pennsylvania State University, University Park, PA 16802, USA

关键词: Glycosylasparaginase;    Post-translational activation;    Active-site mutagenesis;   

DOI  :  10.1016/0014-5793(93)81355-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Lysosomal glycosylasparaginase is encoded as a 36.5 kDa polypeptide that is post-translationally processed to subunits of 19.5 kDa (heavy) and 15 kDa (light). Recombinant glycosylasparaginase has been expressed in Spodoptera frugiperda insect cells enabling the precursor and processed forms to be isolated and their catalytic potential determined. Only the subunit conformation was functional indicating glyeosylasparaginase is encoded as an inactive zymogen. The newly created amino terminal residue of the light subunit following maturation, Thr-206, is believed to be involved in the catalytic mechanism [1992, J. Biol. Chem. 267,6855-6858]. Here we have constructed two amino acid substitution mutants replacing Thr-206 with Ala-206 or Ser-206 and demonstrate that both destroy enzyme activity.

【 授权许可】

Unknown