TRITRICHOMONAS-FETUS - ISOLATION AND CHARACTERIZATION OF THE GOLGI-COMPLEX

An effective methodology to isolate and characterize the Golgi complex of Tritrichomonas foetus is described in this work. Using sucrose den sity gradient centrifugation, two highly enriched Golgi fractions (GF( 1) and GF(2)) wars obtained, Enzymatic assays of GF(1) and GF(2) showe d a strong enrichment in galactosyltransferase activity (20- and 7-fol d, respectively), with minimal contamination with other organelles. Th e GF fraction was further subfractionated by alkaline treatment, which resulted in the production of Golgi content and membrane subfractions . Electron microscopic observations of intact cells or Golgi fractions fixed in solutions containing glutaraldehyde and tannic acid, as weil as of deep-etched replicas of isolated fractions, revealed the presen ce of discrete bridges only between closely apposed cisternae. (C) 199 6 Academic Press, Inc.