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Sulphation of a cell surface glycoprotein from Volvox carteri: Evidence for a membrane-bound sulfokinase working with PAPS

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  • 570 Biowissenschaften
  • Biologie
  • 610 Medizin
  • Biology


PII: 0014-5793(82)80499-7 Volume 139. number 1 FEBS LETTERS March 1982 SULPHATION OF A CELL SURFACE GLYCOPROTEIN FROM VOL VOX CAR TERI Evidence for a membrane-bound sulfokinase working with PAPS Peter WILLADSEN and Manfred SIMPER* CSIRO, Division of Animal Nutrition, Long Pocket Laboratories, Private Bag no. 3, Indooroopilly, QLD 4068, Australia and Wziversit~t Regensburg, Fachbereich Biologic und Vorkl. Medizin, Institut fi2r Biochemie, Genetik und Mikrobiologie, 8400 Regensburg, FRG Received 25 January 1982 1. Introduction Sulphated glycoproteins have been implicated in the control of cell growth and differentiation: fibro- nectins [ 1,2]; glycoproteins in developing sea urchin embryos [3]; and a surface glycoprotein from V&ox curteri [4]. This V&ox glycoprotein, of 185 000 app. M, [4] appears to be involved in the control of the differentiation of the organism during embryogenesis. Although PAPS or APS must be involved as sulphate donor in the synthesis of such molecules, the incor- poration of sulphate from PAPS into a sulphated glycoprotein has been demonstrated only in [5]. It was therefore of interest to partially characterize the sulphate-transfer system operative in Volvos. 2. Materials and methods The growth of Volvox carterif. nagariensis female strain HKlO, pulse labelling of Vobox in vivo with [35S]sulphate, SDS-polyacrylamide gel electropho- resis on 6% gels, fluorography and the preparation of Volvox sonicates were as in [4,6]. During sonication, gonidia were rapidly destroyed and, although a few somatic cells retained their shape, they were perme- able to trypan blue. Membrane and particulate mate- rial was isolated by centrifugation of 1 ml samples of sonicate on 4 ml cushions of 2% sucrose at 110 000 X g for 70 min. Abbreviations: APS, adenosine 5’-phosphosulphate; PAPS, 3’-phosphoadenosine 5’-phosphosulphate 2.1. Preparation of PAPS and APS PAPS was synthesized using the yeast system [7] and isolated by chromatogra

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