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beta]-Glucan Synthesis in the Cotton Fiber (IV. In Vitro Assembly of the Cellulose I Allomorph)


, : beta]-Glucan Synthesis in the Cotton Fiber (IV. In Vitro Assembly of the Cellulose I Allomorph). Plant Physiology 107(1): 111-123

In vitro assembly of cellulose from plasma membrane extracts of the cotton (Gossypium hirsutum) fiber was enriched by a combination of 3-(N-morpholino)propanesulfonic acid extraction buffer and two independent digitonin solubilization steps consisting of 0.05% digitonin (SE1) followed by 1% digitonin (SE2). Glucan synthase activity assays revealed that, although the SE2 fraction possessed higher activity, only 8.6% of the in vitro product survived acetic/nitric acid treatment. On the other hand, the SE1 fraction was less active, but 32.1% of the total glucan in vitro product was resistant to acetic/nitric acid. In vitro products synthesized from the SE1 fraction contained [beta]-1,3-glucan and fibrillar cellulose I, whereas the SE2 fraction produced [beta]-1,3-glucan and cellulose II. Both celluloses assembled in vitro were labeled with cellobiohydrolase I-gold complex, and the electron diffraction patterns of both products from SE1 and SE2 revealed cellulose I and cellulose II, respectively. Contamination of native cellulose was ruled out by extensive evidence from autoradiography of the ethanol-insoluble and acetic/nitric acid-insoluble materials, including three different controls.

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Accession: 047994582

PMID: 12228346

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Kudlicka, K.; Brown, R.M.J.; Li, L.K.; Lee, J.H.; Shin, H.; Kuga, S., 1995: Beta -Glucan synthesis in the cotton fiber. IV. In vitro assembly of the cellulose I allomorph. In vitro assembly of cellulose from plasma membrane extracts of cotton fibre was enriched by a combination of 3-(N-morpholino)propanesulfonic acid extraction buffer and 2 independent digitonin solubilization steps consisting of 0.05% digitonin (SE...

Kudlicka, K.; Brown, R.M.lcolm,. Jr.;; Li, L.; Lee, J., H.; Shin, H.; Kuga, S., 1995: Beta-Glucan synthesis in the cotton fiber. IV. In vitro assembly of the cellulose 1 allomorph. In vitro assembly of cellulose from plasma membrane extracts of the cotton (Gossypium hirsutum) fiber was enriched by a combination of 3-(N-morpholino)propanesulfonic acid extraction buffer and two independent digitonin solubilization steps consis...

Kudlicka, K.; Brown, R.Malcolm, J.; Li, L., 1995: B-glucan synthesis in the cotton fiber. IV. In vitro assembly of the cellulose I allomorph. Plant Physiology 107: 1-23

Okuda, K.L., L.K.dlicka, K.K.ga, S.B.own, R.J., 1993: Beta-Glucan synthesis in the cotton fiber. I. Identification of beta-1,4- and beta-1,3-glucan synthesized in vitro. In vitro beta-glucan products were synthesized by digitonin-solubilized enzyme preparations from plasma membrane-enriched fractions of cotton (Gossypium hirsutum) fiber cells. The reaction mixture favoring beta-1,4-glucan synthesis included the fo...

Okuda, K.; Li, L.; Kudlicka, K.; Kuga, S.; Brown, R.M., 1993: beta]-Glucan Synthesis in the Cotton Fiber (I. Identification of [beta]-1,4- and [beta]-1,3-Glucans Synthesized in Vitro). In vitro [beta]-glucan products were synthesized by digitonin-solubilized enzyme preparations from plasma membrane-enriched fractions of cotton (Gossypium hirsutum) fiber cells. The reaction mixture favoring [beta]-1,4-glucan synthesis included th...

Li, L.; Brown, R.M., 1993: beta]-Glucan Synthesis in the Cotton Fiber (II. Regulation and Kinetic Properties of [beta]-Glucan Synthases. The regulation and kinetic properties of cellulose synthase as well as [beta]-1,3-glucan synthase have been studied. The cellulose was detected using acetic/nitric acid insolubility as an indicator of cellulose (this product contained only [beta]-...

Li, L.; Drake, R.R.; Clement, S.; Brown, R.M., 1993: beta]-Glucan Synthesis in the Cotton Fiber (III. Identification of UDP-Glucose-Binding Subunits of [beta]-Glucan Synthases by Photoaffinity Labeling with [[beta]-32P]5[prime]-N3-UDP-Glucose. Using differential product entrapment and photolabeling under specifying conditions, we identifIed a 37-kD polypeptide as the best candidate among the UDP-glucose-binding polypeptides for the catalytic subunit of cotton (Gossypium hirsutum) cellul...

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Kudlicka, K.B.own, R.J.; Li, L.S.in, H.K.ga, S., 1995: Beta-Glucan synthesis in the cotton fiber. In vitro assembly of cellulose from plasma membrane extracts of the cotton (Gossypium hirsutum) fiber was enriched by a combination of 3-(N-morpholino)propanesulfonic acid extraction buffer and two independent digitonin solubilization steps consis...