Biosynthesis of the Fungal Wall-Mechanisms and Implications The First Fleming Lecture

Gooday, Graham W.

doi:10.1099/00221287-99-1-1

Cited by 122 publications

(53 citation statements)

References 29 publications

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“…Electron microscopy and high-resolution autoradiography of regenerating protoplasts of Schizophyllum commune (van der Valk & Wessels, 1976, 1977 have shown that chitin is only synthesized in close association with the plasma membrane. The present study was undertaken to obtain more information about the localization and activity of chitin synthase in subcellular fractions of these protoplasts.…”

Section: A V E R M E U L E N M B J M R a E V E N A N D J Gmentioning

confidence: 99%

Localization of Chitin Synthase Activity in Subcellular Fractions of Schizophyllum commune Protoplasts

Vermeulen

Raeven

Wessels

1979

Journal of General Microbiology

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Plasma membranes of Schizophyllum commune were stabilized against fragmentation by coating protoplasts with concanavalin A (Con A). A uniform distribution of Con A over the membrane was demonstrated cytochemically. After lysis of the protoplasts in the presence of an inhibitor of proteolysis, plasma membranes were purified and, together with other subcellular fractions, assayed for chitin synthase activity. About 50% of the chitin synthase activity was associated with the plasma membranes and largely occurred in an active form. About 30% of the chitin synthase, in an inactive form, was recovered in fractions sedimenting at 40000 to 300000 g. The product of the plasma membrane-bound enzyme was shown to be a-chitin occurring as microfibrils (about 6 nm wide). The chitin synthase could not be detached from the plasma membranes by incubation with substrate and activator at 0 "C and gradient centrifugation showed that the synthesized chitin remained associated with the plasma membranes.

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Section: A V E R M E U L E N M B J M R a E V E N A N D J Gmentioning

confidence: 99%

Localization of Chitin Synthase Activity in Subcellular Fractions of Schizophyllum commune Protoplasts

Vermeulen

Raeven

Wessels

1979

Journal of General Microbiology

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“…An apparent K; of 0 ·04 flM was calculated for the polyoxin-D inhibition (assuming competitive inhibition). This is)ower than the Ki values determined for polyoxin-D with the fungal chitin synthases (Gooday 1977). …”

Section: Polyoxin-dmentioning

confidence: 63%

“…This structural analogue of UDPAG, which is a competitive inhibitor of the fungal chitin synthases (Gooday 1977), inhibited incorporation by the homogenates when added at low concentrations. However, as shown in Fig.…”

Section: Polyoxin-dmentioning

confidence: 98%

“…This enzyme has been studied in fungi and yeasts (Gooday 1977) and, recently, in cell-free membrane and microsomal preparations of extracts of whole insects (Cohen and Casida 1980a;Mayer et al 1980). The tissue source from which these insect activities derive has not been established but Cohen and Casida (1980a) suggest that in their preparations from Tribolium infestans the enzyme comes from the gut, where it is involved in the synthesis of peri trophic membranes.…”

Section: Introductionmentioning

confidence: 99%

“…One group of fungicides which inhibit chitin deposition are the polyoxins (Misato et al 1979). These compounds are structural analogues of UDP-N-acetylglucosamine and have been shown to competitively inhibit fungal chitin synthases (Gooday 1977). The benzoylphenylurea insecticides also inhibit cuticular chitin deposition in a variety of insects (Sowa and Marks 1975;Hajjar and Casida 1978) and appear to act on the final polymerization step of the chitin biosynthetic pathway (Verloop and Ferrell 1977), but it is still unclear whether they directly inhibit chitin synthase.…”

Section: Introductionmentioning

confidence: 99%

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Integumental Chitin Synthase Activity in Cell-free Extracts of Larvae of the Australian Sheep Blowfly, Lucilia cuprina, and Two Other Species of Diptera

Turnbull¹,

Howells²

1983

Aust. Jnl. Of Bio. Sci.

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Chitin synthase activity has been demonstrated in crude homogenates of larval integuments from L. cuprina and in similar preparations from Musca domestica and Calliphora erythrocephala. This is the first report of an insect integumental chitin synthase. This activity brings about the incorporation of radioactivity from UDP-N-acetyl-[14C]glucosamine into an ethanol-and alkali-insoluble form. A major part of this labelled product has been characterized as chitin by its insolubility in alkali, resistance to degradation by proteases and its susceptibility to digestion by chitinase and RCI. Most of the radioactivity solubilized during digestion by chitinase co-migrates with N-acetylglucosamine, glucosamine and chitobiose during paper chromatography. Some radioactivity also becomes incorporated into non-chitin products in this system. There is substantial evidence that incorporation is not brought about by whole epidermal cells or by microbial contamination in the homogenates. The extent of incorporation obtained with the homogenates is limited by the presence of degradative enzymes which rapidly break down the substrate (UDP-N-acetylglucosamine). The incorporation was partially inhibited (50-70%) by both polyoxin-D (apparent Ki 0'04I1M) and diftubenzuron (apparent . This is the first report of a cell-free chitin-synthesizing system derived from insect tissue which is sensitive to inhibition by diftubenzuron.

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The Synthesis and Degradation of Chitin

Cabib

1987

Advances in Enzymology - And Related Areas of Molecular Biology

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Biosynthesis of the Fungal Wall-Mechanisms and Implications The First Fleming Lecture

Cited by 122 publications

References 29 publications

Localization of Chitin Synthase Activity in Subcellular Fractions of Schizophyllum commune Protoplasts

Localization of Chitin Synthase Activity in Subcellular Fractions of Schizophyllum commune Protoplasts

Integumental Chitin Synthase Activity in Cell-free Extracts of Larvae of the Australian Sheep Blowfly, Lucilia cuprina, and Two Other Species of Diptera

The Synthesis and Degradation of Chitin

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