Examination of ribosome-like particles in isolated prolamellar bodies

Potential methods for the preparation of fractions enriched in prolamellar bodies (PLBs) were examined in detail. Sucrose density gradient centrifugation methods gave fractions consisting almost exclusively of PLBs whilst those methods employing differential centrifugation were quite successful but contained greater quantities of lamellar membranes. Greater difficulty was experienced in obtaining detached PLBs which retained their ribosome-like lattice particles. No modification to density gradient procedures was found which retained these particles but the omission of ethylene diaminetetraacetic acid (EDTA) from all media including that of lysis gave a hint that this was possible with differential centrifugal methods. This was developed to produce a successful method for the preparation of PLBs which retain the ribosome-like particles of the lattice. Such fractions from Avena sativa L. and Hordeum vulgare L. were treated with ribonuclease which completely removed these particles from the lattice structures implying that they may be ribosomal in nature. EDTA apparently has a critical effect on PLB structure at concentration lower than those that effect the chloroplast coupling factor particles but it is not known if it is a direct effort of PLB membranes, on the lattice particles or both.Abbreviations PLB prolamellar body - EDTA Ethylene diaminetetra-acetic acid - MOPS morpholinopropane sulphonic acid - CF₁ chloroplast coupling factor particles - SDS sodium dodecyl sulphate