Purification and characterization of 16 S chitin synthetase particles from cell walls ofMucor rouxii

Lending, Craig R.; Leal-Morales, Carlos A.; Flores-Martínez, Alberto; Bracker, Charles E.; Bartnicki-García, S.

doi:10.1016/0147-5975(91)90003-v

Cited by 22 publications

(14 citation statements)

References 9 publications

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“…However some explanation may be needed for the 8.2-kb mRNA in class IV. Biochemical studies have shown that a highly purified CHS function from Mucor rouxii, which is a typical Zygomycetes, is associated with four polypeptides (Lending et al, 1991). The long mRNA may provide a mechanism for encoding more than one protein subunit required for CHS activity.…”

Section: Discussionmentioning

confidence: 99%

Multiple genes for chitin synthase in the zygomycete fungus Phycomyces blakesleeanus.

Miyazaki

Ootaki

1997

J. Gen. Appl. Microbiol.

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Section: Discussionmentioning

confidence: 99%

Multiple genes for chitin synthase in the zygomycete fungus Phycomyces blakesleeanus.

Miyazaki

Ootaki

1997

J. Gen. Appl. Microbiol.

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“…The assay mixture was incubated at 25 "C for 30 min and the reaction was stopped with a drop of glacial acetic acid. Membrane-bound glucan synthase (50 pl) was assayed with 4 mM cellobiose, 0 3 mM dithiothreitol (DTT), 0-8 mM UDP-Glc and 4 nCi UDP- Samples were treated with sample buffer as described by Lending et al (1991). Electrophoresis was performed overnight at a constant current of 10 mA.…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, it is not possible to state unequivocally which of the detected polypeptides belong to the chitin synthase complex. A similar study of chitin synthase from M. rouxii solubilized with digitonin (16 S particles) revealed several polypeptide bands in the final step of purification, four of which (21, 23, 33 and 39 kDa) were proposed to be the components of a macromolecular complex that synthesizes chitin microfibrils (Lending et al, 1991). That S. monoica and M. rouxii yielded a different mix of polypeptides is not surprising given the different size of the solubilized enzyme from S. monoica, its lack of a net cationic charge, and the sharp biochemical differences between Oomycetes and Zygomycetes (Bartnicki-Garcia, 1970).…”

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confidence: 99%

“…So far, however, no chitin synthase polypeptide(s) of any organism has been unequivocally purified to homogeneity. Moreover, it appears that fungal chitin synthase does not consist of a single polypeptide but a multipeptide complex (Lending et al, 1991). Thus, the smallest units of active chitin synthase ever isolated are the so-called 16 S complexes (Ruiz-Herrera et af., 1980;Lending et af., 1991).…”

Section: Introductionmentioning

confidence: 99%

“…The enzyme from S. monoica was not stimulated much by any of several phospholipids tested, contrary to the behaviour of chitin synthases from Saccharomyces cerevisiae (Duran & Cabib, 1978) ; Schizophyllum commune (Vermeulen & Wessels, 1983) ; Candida albicans (Braun & Calderone, 1979); M . rouxii (Lending et al, 1991) ; Coprinus cinereus (Montgomery & Gooday, 1985) or Fusarium graminearum (Binks et al, 1990). 4.…”

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confidence: 99%

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The properties and localization of Saprolegnia monoica chitin synthase differ from those of other fungi

1997

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a n d Sa I om 6 n B a r t n i c k i -G a rc fa3 The presence of non-fibrillar a-chitin in cellulosic fungi (class Oomycetes) poses intriguing questions as to i t s role, subcellular localization and evolutionary significance. Previous studies reported on the similarity of chitin synthase from Saprolegnia monoica with that of other fungi. The present work describes important dissimilarities. There was no evidence that the chitin synthase of S. monoica was present in small low-density vesicles (chitosomes). Chitin synthase sedimented with membranous components of high specific gravity (sp. gr. 1-177) that could be partially but distinctly separated from membranes harbouring most of the 1,3-&glucan synthase in the cell (sp. gr. 1.158).In contrast to other fungi, the chitin synthase from 5. monoica was greatly stimulated by digitonin: both membrane-bound and dissociated chitin synthase showed little activity in the absence of digitonin. As in other fungi, the chitin synthase from S. monoica was solubilized by digitonin and remained zymogenic after dissociation. However, unlike the enzyme from other fungi, the solubilized chitin synthase of S. monoica had a lower sedimentation coefficient, was not stimulated by phospholipids and was not inhibited by high concentrations of digitonin. Unlike the enzyme from Mucor rouxii, the solubilized chitin synthase from 5. monoica did not bind to a cation exchanger. The enzyme was partially purified by a four-step scheme that included sucrose density-gradient centrifugation, a single passage through a strong anion exchanger and two consecutive passages through a weak anion exchanger. The final preparation contained five to seven polypeptide bands that cochromatographed with the chitin synthase activity, some of which may be part of a presumed chitin synthase macromolecular complex.

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Biochemistry of chitin synthase

Merz

Horsch²,

Nyhlén³

et al. 1999

Chitin and Chitinases

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Purification and characterization of 16 S chitin synthetase particles from cell walls ofMucor rouxii

Cited by 22 publications

References 9 publications

Multiple genes for chitin synthase in the zygomycete fungus Phycomyces blakesleeanus.

Multiple genes for chitin synthase in the zygomycete fungus Phycomyces blakesleeanus.

The properties and localization of Saprolegnia monoica chitin synthase differ from those of other fungi

Biochemistry of chitin synthase

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