NDLI: Molecular cloning and biochemical characterization of medaka (Oryzias latipes) lysosomal neu4 sialidase

Content Provider	Springer Nature Link
Author	Shiozaki, Kazuhiro Ryuzo, Sena Matsushita, Naoto Ikeda, Asami Takeshita, Kazuki Chigwechokha, Petros Kingstone Komatsu, Masaharu Miyagi, Taeko
Copyright Year	2014
Abstract	Glycoconjugates are known to be involved in many physiological events in vertebrates. Sialidase is one of the glycosidases, which removes sialic acid from glycoconjugates. In mammals, the properties and physiological functions of sialidases have been investigated, while there is little understanding of fish sialidase. Here, to investigate the significance of fish neu4 sialidase, neu4 gene was cloned from medaka brain mRNA and identified. Sialidase-specific motifs (GPG, YRVP and Asp-Box) were well conserved in the medaka neu4 polypeptide. Optimal pH of medaka neu4 sialidase was 4.6, but its activity was sustained even at neutral and weak alkaline pH. The neu4 considerably cleaved sialic acid from 4-methylumbelliferyl-N-acetyl-α-d-neuraminic acid and sialyllactose, but not from ganglioside and fetuin, which are good substrates for human NEU4. neu4 activity was mostly detected in mitochondria/lysosome fraction after biochemical fractionation, and indirect immunofluorescence assays revealed neu4 localization in lysosome in neu4 overexpressed cells. Next, developmental change in medaka neu4 and other sialidase mRNA levels were estimated by real-time PCR. Each sialidases showed different expression patterns in embryonic development: neu4 was up-regulated at late developmental stage in embryo, and neu3a mRNA level was quite high in 0.5 dpf. On the other hand, neu3b expression was drastically increased after hatching, suggesting that each sialidase may play a different role in embryonic development.
Starting Page	1461
Ending Page	1472
Page Count	12
File Format	PDF
ISSN	09201742
Journal	Fish Physiology and Biochemistry
Volume Number	40
Issue Number	5
e-ISSN	15735168
Language	English
Publisher	Springer Netherlands
Publisher Date	2014-04-18
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Development Embryo Glycoconjugates Japanese medaka Neu4 Sialidase Freshwater & Marine Ecology Animal Physiology Animal Anatomy / Morphology / Histology Animal Biochemistry Zoology
Content Type	Text
Resource Type	Article
Subject	Physiology Medicine Biochemistry Aquatic Science

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