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Subcellular localization of mannosyl transferase and glycoprotein biosynthesis in castor bean endosperm

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Abstract

Differential and sucrose density gradient centrifugation have shown that the mannosyl transferase present in germinating castor bean endosperm cells which catalyses the synthesis of mannosyl-phosphoryl-polyisoprenol is exclusively located in the endoplasmic reticulum membrane. This intracellular location was confirmed using both ribosome-denuded microsomes isolated in the presence of EDTA and rough-surfaced microsomes isolated in the presence of excess Mg2+ added to maintain ribosome-membrane attachment. Separation of organelles following the incubation of crude particulate fractions with GDP[14C]mannose demonstrated that most of the mannolipid thus formed remained associated with the microsomal fraction. When organelles were isolated from intact tissue which had previously been incubated with GDP[14C]mannose, [14C]glycoprotein was found to be associated with other cellular fractions in addition to the microsomes, in particular the glyoxysomes. The kinetics of radioactive labelling of these organelles suggest that [14C]glycoprotein appears initially in the microsomal fraction and subsequently accumulates in the glyoxysomes. Subfractionation of isolated, [14C]glycoprotein-labelled glyoxysomes established that over 80% of the total radioactivity was present in the membrane, while sodium dodecyl sulphate-polyacrylamide gel electrophoresis of solubilized glyoxysomal membranes showed that the [14C]sugar moiety was associated with several, but not all, constituent polypeptides.

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Abbreviations

ER:

endoplasmic reticulum

TCA:

trichloroacetic acid

SDS:

sodium dodecylsulphate

GDP:

guanosine diphosphate

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Authors and Affiliations

  1. Postgraduate School of Biological Sciences, University of Bradford, BD7 IDP, Bradford, U.K.

    R. B. Mellor & J. M. Lord

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  1. R. B. Mellor
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  2. J. M. Lord
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Mellor, R.B., Lord, J.M. Subcellular localization of mannosyl transferase and glycoprotein biosynthesis in castor bean endosperm. Planta 146, 147–153 (1979). https://doi.org/10.1007/BF00388225

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  • DOI: https://doi.org/10.1007/BF00388225

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