Expression of lysosomal protective protein/cathepsin A in a stably transformed human neuroblastoma cell line during bi-directional differentiation into neuronal and Schwannian cells

Itoh, Kohji; Satoh, Y.; Kadota, Yoshito; Oheda, Yukako; Kuwahara, Jun; Shimmoto, Michie; Sakuraba, Hitoshi

doi:10.1016/j.neuint.2003.07.001

Cited by 4 publications

(1 citation statement)

References 37 publications

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“…Each fraction, including pass-through (PT), washing, and eluate, obtained by Ni-column chromatography of the conditioned medium of CHO/CTSA-6His containing the proCTSA-6His was applied to SDS-PAGE under reducing conditions and then electroblotted onto polyvinylidene difluoride membranes (Immobilon; Nippon Millipore, Tokyo, Japan). 43 The blotted membranes were blocked with 50% Blocking One (Nacalai tesque, Kyoto, Japan) in Tris-buffered saline (TBS), followed by incubation with anti-CTSA or anti-His antibody and HRP-conjugated anti-rabbit IgG or HRP-conjugated anti-mouse IgG. The antibody-reacting bands were visualized with a chemiluminescence detection system (Western Lightning Chemiluminescence Reagent PLUS; PerkinElmer Life Sciences, Inc., Boston, MA) according to the manufacturer's protocol.…”

Section: Western Blottingmentioning

confidence: 99%

Reversal of neuroinflammation in novel GS model mice by single i.c.v. administration of CHO-derived rhCTSA precursor protein

Horii

Iniwa

Onitsuka

et al. 2022

Molecular Therapy - Methods & Clinical Development

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Section: Western Blottingmentioning

confidence: 99%

Reversal of neuroinflammation in novel GS model mice by single i.c.v. administration of CHO-derived rhCTSA precursor protein

Horii

Iniwa

Onitsuka

et al. 2022

Molecular Therapy - Methods & Clinical Development

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Cathepsin A protein from the accessory sex gland of the Chinese mitten crab (Eriocheir sinensis) plays a key role in spermatophore digestion

Wang¹,

Fang²,

Wang³

et al. 2013

Journal of Insect Physiology

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Elimination of abnormal sialylglycoproteins in fibroblasts with sialidosis and galactosialidosis by normal gene transfer and enzyme replacement

Oheda¹,

Kotani²,

Murata³

et al. 2005

Glycobiology

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Sialidosis and galactosialidosis are lysosomal storage diseases caused by the genetic defects of lysosomal sialidase (neuraminidase-1; NEU1) and lysosomal protective protein/cathepsin A (PPCA), respectively, associated with a NEU1 deficiency, excessive accumulation of sialylglycoconjugates, and development of progressive neurosomatic manifestations; in addition, the latter disorder is accompanied by simultaneous deficiencies of beta-galactosidase and cathepsin A. We demonstrated that a few soluble N-glycosylated proteins carrying sialyloligosaccharides sensitive to glycopeptidase F (GPF) can be specifically detected in cultured fibroblasts from sialidosis and galactosialidosis cases by blotting with a Maackia amurensis (MAM) lectin. We also examined the therapeutic effects of normal gene transfer and enzyme replacement by evaluating the decreases in sialylglycoconjugates accumulated in fibroblasts with these NEU1 deficiencies. The specific N-glycosylated proteins detected on MAM lectin blotting as well as the granular lysosomal fluorescence due to an avidin-FITC/biotinylated MAM lectin conjugate in sialidosis and galactosialidosis fibroblasts disappeared in parallel with the restoration of the intracellular NEU1 activity after transfection of the recombinant NEU1 fused to HA tag sequence and the wild-type PPCA cDNA as well as administration of the recombinant PPCA precursor protein. The detection method for the abnormal sialylglycoproteins in cultured cells involving MAM lectin was demonstrated to be useful not only for biochemical and diagnostic analyses of NEU1 deficiencies but also for therapeutic evaluation of these conditions.

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Expression of lysosomal protective protein/cathepsin A in a stably transformed human neuroblastoma cell line during bi-directional differentiation into neuronal and Schwannian cells

Cited by 4 publications

References 37 publications

Reversal of neuroinflammation in novel GS model mice by single i.c.v. administration of CHO-derived rhCTSA precursor protein

Reversal of neuroinflammation in novel GS model mice by single i.c.v. administration of CHO-derived rhCTSA precursor protein

Cathepsin A protein from the accessory sex gland of the Chinese mitten crab (Eriocheir sinensis) plays a key role in spermatophore digestion

Elimination of abnormal sialylglycoproteins in fibroblasts with sialidosis and galactosialidosis by normal gene transfer and enzyme replacement

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