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A 1,4-β-D-glucan-synthase system fromDictyostelium discoideum

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Abstract

Particulate membrane preparations have been isolated from culminatingDictyostelium discoideum cells. The preparations incorporated glucose from uridine 5′-diphosphate-glucose into a glucose polymer or polymers. These have been shown to be homopolymers ofβ-linked glucose. A high percentage (78% by methylation analysis) of the linkages formed are 1,4-linkages and a lower percentage (12%) are 1,3-linkages. The glucan-synthase complex present in the particulate membrane preparation has an apparent Km of 0.28 mM and a Vmax of 1.59 nmol·min−1·(mg protein)−1. The enzyme system is dependent upon Mg2+ and cellobiose for maximal activity, but is inhibited by millimolar levels of Ca2+. Particulate membrane preparations were made from cells at various times during a synchronous developmental time course and demonstrated that the glucan-synthase activity appeared at the tight-aggregate stage of development.

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Abbreviations

EDTA:

ethylenediaminetetraacetic acid

EGTA:

ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid

Hepes:

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

UDP:

uridine 5′-diphosphate

UDPGlc:

UDP-glucose

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Author notes

  1. R. L. Blanton

    Present address: Department of Biological Sciences, Texas Tech University, P.O. Box 4149, 79409-3131, Lubbock, TX, USA

Authors and Affiliations

  1. Department of Biochemistry, University of Cambridge, Tennis Court Road, CB2 1QW, Cambridge, UK

    R. L. Blanton & D. H. Northcote

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  1. R. L. Blanton
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Additional information

The authors thank Dr. R.R. Kay for provision of cultures and advice, Dr. P. Rubery for the loan of an environmental shaker, Mr. R. Davy for performing the methylation and gas-liquid chromatography analyses, Mr. M. Grimson for the sectioned vesicles and the shadowed stalk preparation, Dr. R. Shaw for provision of the non-linear regression computer program, and Dr. C.H. Haigler for helpful discussions and critical reading of the manuscript. R.L.B. also wishes to thank Messrs. R. Smyer and B. Harris for technical assistance. This paper is based in part upon research supported by the Texas Advanced Research Program under Grant No. 1388.

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Blanton, R.L., Northcote, D.H. A 1,4-β-D-glucan-synthase system fromDictyostelium discoideum . Planta 180, 324–332 (1990). https://doi.org/10.1007/BF01160387

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