Abstract
Sealed membrane vesicles were prepared from microsomes and glyoxysomes isolated from the endosperm tissue of germinating castor bean. Peripheral-membrane proteins together with soluble protein present in the luminal space of the microsomes or the matrix of the glyoxysomes were released from intact organelles by osmotic shock in the presence of salt. The washed membrane vesicles were linked to cyanogen-bromide-activated Sepharose. Where appropriate, the immobilized vesicles were made permeable to protein molecules by controlled detergent treatment which did not result in significant solubilization of the lipid bilayer. Released luminal proteins were allowed to interact with the membrane vesicles under conditions which gave them access to the cytoplasmic surface only or to both the cytoplasmic and luminal surfaces. While microsomal luminal proteins did not interact with either surface of the membrane vesicles, glyoxysomal matrix proteins specifically bound to the luminal surface of the glyoxysomal membrane. Binding seemed to be effected via the oligosaccharide chains of glyoxysomal membrane glycoproteins since (a) bound proteins could be released by elution with sugar solution, and (b) solubilized glyoxysomal membrane proteins specifically interacted with immobilized lectins.
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Abbreviations
- CDP-choline:
-
cytidine diphosphocholine
- ER:
-
endoplasmic reticulum
References
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Harson, M.M., Conder, M.J. & Lord, J.M. Endoplasmic reticulum and glyoxysomal membranes from castor-bean endosperm: interaction between membrane glycoproteins and organelle matrix proteins. Planta 157, 143–149 (1983). https://doi.org/10.1007/BF00393648
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DOI: https://doi.org/10.1007/BF00393648