Fluorogenic sialic acid glycosides for quantification of sialidase activity upon unnatural substrates

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS

卷:23

Article

Zamora, Cristina Y.¹  d'Alarcao, Marc²  Kumar, Krishna^1,3 

[1] Tufts Univ, Dept Chem, Medford, MA 02155 USA

[2] San Jose State Univ, Dept Chem, San Jose, CA 95192 USA

[3] Tufts Med Ctr, Ctr Canc, Boston, MA 02110 USA

关键词: Sialidases;    Neuraminidases;    4-MU;    Sialic acid derivatives;    Glycoengineering;    Fluorination;    NEU1;    NEU2;    NEU3;   

DOI  :  10.1016/j.bmcl.2013.03.076

来源: Elsevier

【 摘 要 】

Herein we report the synthesis of N-acetyl neuraminic acid derivatives as 4-methylumbelliferyl glycosides and their use in fluorometrically quantifying human and bacterial sialidase activity and substrate specificities. We found that sialidases in the human promyelocytic leukemic cell line HL60 were able to cleave sialic acid substrates with fluorinated C-5 modifications, in some cases to a greater degree than the natural N-acetyl functionality. Human sialidases isoforms were also able to cleave unnatural substrates with bulky and hydrophobic C-5 modifications. In contrast, we found that a bacterial sialidase isolated from Clostridium perfringens to be less tolerant of sialic acid derivatization at this position, with virtually no cleavage of these glycosides observed. From our results, we conclude that human sialidase activity is a significant factor in sialic acid metabolic glycoengineering efforts utilizing unnatural sialic acid derivatives. Our fluorogenic probes have enabled further understanding of the activities and substrate specificities of human sialidases in a cellular context. (C) 2013 Elsevier Ltd. All rights reserved.

【 授权许可】

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