Purification and characterization of β-N-acetylhexosaminidase from maize seedlings

Oikawa, Akira; Itoh, Eriko; Ishihara, Atsushi; Iwamura, Hajime

doi:10.1078/0176-1617-01089

Cited by 23 publications

(15 citation statements)

References 39 publications

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“…A b-hexosaminidase isolated from cotton (Gossypium hirsutum) seedlings was found to release GlcNAc residues from glycoprotein substrates (Yi, 1981). However, only two of the four isoforms from fenugreek were able to remove b1,2-linked GlcNAc residues from N-glycans (Bouquelet and Spik, 1978), while the purified b-N-acetylhexosaminidase from maize (Zea mays) seedlings displayed no detectable activity with N-glycan and glycoprotein substrates (Oikawa et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

“…in seeds, leaves, and suspension cultured cells (Li and Li, 1970;Boller and Kende, 1979;Barber and Ride, 1989;Oikawa et al, 2003). In particular, high levels of b-N-acetylhexosaminidase activity have been detected in germinating seeds (Bouquelet and Spik, 1978;Yi, 1981;Oikawa et al, 2003), suggesting a physiological role in the metabolism of storage glycoproteins (Harris and Chrispeels, 1975).…”

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confidence: 99%

“…In particular, high levels of b-N-acetylhexosaminidase activity have been detected in germinating seeds (Bouquelet and Spik, 1978;Yi, 1981;Oikawa et al, 2003), suggesting a physiological role in the metabolism of storage glycoproteins (Harris and Chrispeels, 1975). A function in defense-related processes has also been proposed since several of the purified b-N-acetylhexosaminidases could degrade chitin oligomers (Li and Li, 1970;Yi, 1981;Oikawa et al, 2003). However, no plant b-hexosaminidase has yet been characterized on the molecular level.…”

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Enzymatic Properties and Subcellular Localization of Arabidopsis β-N-Acetylhexosaminidases

et al. 2007

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Plant glycoproteins contain substantial amounts of paucimannosidic N-glycans lacking terminal GlcNAc residues at their nonreducing ends. It has been proposed that this is due to the action of b-hexosaminidases during late stages of N-glycan processing or in the course of N-glycan turnover. We have now cloned the three putative b-hexosaminidase sequences present in the Arabidopsis (Arabidopsis thaliana) genome. When heterologously expressed as soluble forms in Spodoptera frugiperda cells, the enzymes (termed HEXO1-3) could all hydrolyze the synthetic substrates p-nitrophenyl-2-acetamido-2-deoxy-b-Dglucopyranoside, p-nitrophenyl-2-acetamido-2-deoxy-b-D-galactopyranoside, 4-methylumbelliferyl-2-acetamido-2-deoxy-b-Dglucopyranoside, and 4-methylumbelliferyl-6-sulfo-2-acetamido-2-deoxy-b-D-glucopyranoside, albeit to a varying extent. HEXO1 to HEXO3 were further able to degrade pyridylaminated chitotriose, whereas pyridylaminated chitobiose was only cleaved by HEXO1. With N-glycan substrates, HEXO1 displayed a much higher specific activity than HEXO2 and HEXO3. Nevertheless, all three enzymes were capable of removing terminal GlcNAc residues from the a1,3-and a1,6-mannosyl branches of biantennary N-glycans without any strict branch preference. Subcellular localization studies with HEXOfluorescent protein fusions transiently expressed in Nicotiana benthamiana plants showed that HEXO1 is a vacuolar protein. In contrast, HEXO2 and HEXO3 are mainly located at the plasma membrane. These results indicate that HEXO1 participates in N-glycan trimming in the vacuole, whereas HEXO2 and/or HEXO3 could be responsible for the processing of N-glycans present on secretory glycoproteins.

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Section: Discussionmentioning

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Enzymatic Properties and Subcellular Localization of Arabidopsis β-N-Acetylhexosaminidases

et al. 2007

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“…In the past only three diVerent exochitinase activities were detected in maize (Oikawa et al 2003). However, four diVerent genes exist in the maize genome.…”

Section: Discussionmentioning

confidence: 99%

Genome-wide identification, expression and chromosomal location of the genes encoding chitinolytic enzymes in Zea mays

Shoresh

Harman

2008

Mol Genet Genomics

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Chitinolytic enzymes are important pathogenesis and stress related proteins. We identiWed 27 putative genes encoding endochitinases in the maize genome via in silico techniques and four exochitinases. Only seven of the endochitinases and segments of the exochitinases were heretofore known. The endochitinases included members of family 19 chitinases (classes I-IV of PR3, II of PR4) and members of family 18 chitinases (class III of PR8). Some similar enzymes were detected on adjacent regions of the same chromosome, and seem to result from duplication events. Most of the genes expressed were identiWed from EST libraries from plants exposed to biotic or abiotic stresses but also from libraries from tissues not exposed to stresses. We isolated proteins from seedlings of maize in the presence or absence of the symbiotic root colonizing fungus Trichoderma harzianum strain T22, and analyzed the activity of chitinolytic enzymes using an in-gel activity assay. The activity bands were identiWed by LC/MS/MS using the database from our in silico study. The identities of the enzymes changed depending on whether or not T22 was present. One activity band of about 95 kDa appeared to be a heterodimer between an exochitinase and any of several diVerent endochitinases. The identity of the endochitinase component appeared to be dependent upon treatment.

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“…The enzyme that catalyses the cleavage of b-N-acetylglucosaminide is known as b-N-acetylglucosaminidase, while an enzyme for b-N-acetylgalactosaminide has not been detected or has not been researched (Oikawa, Itoh, Ishihara, & Iwamura, 2003). In general, b-N-acetylhexosaminidases (b-NAHAs) hydrolyse the b-glycosides of N-acetylglucosaminide and N-acetylgalactosaminide; they are not specific for the aglycone group, although they prefer aryl substituents.…”

Section: Introductionmentioning

confidence: 99%

Characterisation of a β-N-acetylhexosaminidase from a commercial papaya latex preparation

et al. 2011

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Purification and characterization of β-N-acetylhexosaminidase from maize seedlings

Cited by 23 publications

References 39 publications

Enzymatic Properties and Subcellular Localization of Arabidopsis β-N-Acetylhexosaminidases

Enzymatic Properties and Subcellular Localization of Arabidopsis β-N-Acetylhexosaminidases

Genome-wide identification, expression and chromosomal location of the genes encoding chitinolytic enzymes in Zea mays

Characterisation of a β-N-acetylhexosaminidase from a commercial papaya latex preparation

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