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Intracellular membrane structures of the mycoplasma-like organisms inducing carrot yellows.

Intracellular membrane structures from 16 to 22 nm wide were found in the mycoplasma-like organisms causing carrot yellows. These structures are composed of five layers, three electron-dense and two electron-transparent. The membrane structures are situated, as a rule, in the central nucleoid zone of the cells of the mycoplasma-like organisms and may be seen to be in direct contact with the DNA fibrils. Ribosomes may be seen around the membrane structures. In this work we show the morphological connection between the membrane structures and the external elementary membrane of the mycoplasma-like organisms and also invagination of the external membrane into the cell. It is proposed that the intracellular membrane structures arise by means of invagination of the external elementary membrane and form a single system with this. This is the first instance of discovery of intracellular membrane structures in mycoplasmata.

Intracellular Membranes

[Fatty acid composition and unsaturated of intracellular membrane phospholipids from rat liver and hepatoma 27].

The fatty acid composition of phospholipids of mitochondria and microsomes from rat liver and hepatoma 27 was investigated. Basing on the fatty acid and phospholipid composition the unsaturation of the lipid bilayer of the intracellular membranes was calculated. The unsaturation of the phospholipids of the hepatoma mitochondria and microsomes was found to be much lower than that of the corresponding rat liver membranes. The lipid bilayer of the rat liver and hepatoma plasma membranes was shown to be more saturated than that of the intracellular membranes.

Animals

The relative thickness of intracellular membranes in epithelial cells of the ventral lobe of the rat prostate.

The relative thickness of intracellular membranes of epithelial cells in the ventral lobe of the rat prostrate was measured by a densitometric method. Glutaraldehyde perfusion followed by ruthenium tetroxide immersion fixation appeared to be the most suitable method for membrane thickness measurements. By thickness, the membranes could be roughly subdivided into three groups. The inner and outer membranes of the mitochondrion made up the thinnest membranes of the cell. The second group of membranes consisted of the membranes of the rough-surfaced endoplasmic reticulum and the Golgi apparatus, the different faces of the latter organelle, and the Golgi vesicles. The thickest group of membranes included those of the cell membrane, secretory granules, condensing vacuoles, lysosomes, autophagic vacuoles and multivesicular bodies. The differences in thickness of the membranes are probably due to the varying protein/lipid ratio, and the qualities and proportions of the different lipids in the membranes.

Animals

[Intracellular membrane structures of Bdellovibrio bacteriovorus during development in the bacterial host cell].

Electron microscope investigation of intracellular membrane structures of Bd. bacteriovorous during its intracellular growth and development has been carried out. Increased amount of membrane structures has been stated. Along with simply organized invaginations of plasmalemma, complicated membrane structures resembling mesosomes of grampositive bacteria are observed. Localization of these structures testifies to their involvement in the synthesis of exotoxins and exoenzymes.

Bdellovibrio

Vesicular stomatitis virus glycoprotein is anchored to intracellular membranes near its carboxyl end and is proteolytically cleaved at its amino terminus.

The intracellular vesicular stomatitis virus glycoprotein (G) is inserted into membranes such that a small portion of one end of the molecule is exposed on the cytoplasmic surface of the endoplasmic reticulum and is susceptible to proteolytic digestion (T.G. Morrison, C.O. McQuain, and D. Simpson, J. Virol. 28:368-374). We have determined that this region of the G protein contains two methionyl tryptic peptides. The methionyl tryptic peptides of the G protein have been ordered by the use of the antibiotic pactamycin, and the two methionyl tryptic peptides removed by proteolytic digestion of intracellular G protein have been shown to be derived from the carboxyl terminal end of the protein. In addition, we have found that the unglycosylated G protein synthesized in a reticulocyte cell-free reaction migrates on polyacrylamide gels slightly slower than the unglycosylated G protein synthesized in tunicamycin-treated infected cells. We have also compared these G proteins derived from different sources by partial proteolysis (D.W. Cleveland, S.G. Fischer, M.W. Kirschner, and V.K. Laemmli, J. Biol. Chem. 252:1102-1106) and by chymotryptic peptide analysis. We have found minor differences between the two proteins consistent with the removal of 10 to 15 amino acids from the amino terminus of the intracellular G protein.

Cell Line

Use of fluorescence polarization to monitor intracellular membrane changes during temperature acclimation. Correlation with lipid compositional and ultrastructural changes.

Fluorescence polarization of 1,6-diphenylhexatriene (DPH) was used to study the effects of temperature acclimation on Tetrahymena membranes. The physical properties of membrane lipids were found to be highly dependent on cellular growth temperature. DPH polarization in lipids from three different membrane fractions correlated well with earlier freeze-fracture and electron spin resonance observations showing that membrane fluidity progressively decreases in the order microsomes greater than pellicles greater than cilia throughout a wide range of growth temperatures. Changes in membrane lipid fluidity following a shift from high to low growth temperatures proceed rapidly in the microsomes, whereas there is a pronounced lag in the changes of peripheral cell membrane lipids. These data support previous observations that adaptive changes in membrane fluidity proceed via lipid modifications in the endoplasmic reticulum, followed by dissemination of lipid components to other cell membranes. The rapid changes in polarization observed in the microsomal lipids following a temperature shift correspond closely with the time-dependent alterations in both lipid fatty acid composition and freeze-fracture patterns of membrane particle distribution, suggesting that, in the endoplasmic reticulum, lipid phase separation is the primary cause of membrane particle rearrangements.

Animals

Synthesis of intracellular membrane proteins in vitro. Relation between rough endoplasmic reticulum and mitochondrial outer membrane.

Hepatic rough microsomes were incubated in a messenger-dependent protein-synthesizing system from rabbit reticulocytes. Up to 30% of the total product labelled with [35S]methionine, and subsequently recovered with the microsomes, was located in an intrinsic protein fraction associated with these membranes, i.e. was retained by the membrane following extensive sonication in the presence of 1.5 M KCl, 0.1% deoxycholate, and 5 mM ethylenediaminetetra-acetate (EDTA). When products synthesized with the use of membrane-free mRNA from rough microsomes and free polysome were post-incubated with rough microsomes, ribosome-stripped rough microsomes, or outer mitochondrial membrane, low amounts of intrinsic-type polypeptide product were recovered with these membranes. Higher recovery was achieved, however, when ribosome-stripped rough microsomes were added at the beginning of polypeptide synthesis in a reticulocyte lysate supplemented with additional ribosomal-wash factors. Analysis of these products by polyacrylamide gel electrophoresis showed that a number co-migrated with intrinsic proteins located in both rough microsomes and mitochondrial outer membrane. In addition, a prominent in vitro product co-migrated with a major protein which is located in outer mitochondrial membrane fractions, but is barely detectable in rough microsomal fractions. The present experiments were unable to detect a unique set of intrinsic polypeptides which were synthesized and assembled in vitro under the direction of mRNA from free polysomes, and not from rough microsomes. The results suggest that synthesis of at least some intrinsic membrane proteins which are destined for the outer mitochondrial membrane occurs on rough ER in rat liver.

Animals

Effect of intraocular irrigating solutions on intracellular membrane potentials and swelling rate of isolated human and rabbit cornea.

Isolated human and rabbit corneas were incubated in glutathione bicarbonate Ringer solution (GBR), balanced salt solution (BSS), or 0.9% NaCl solution. The swelling rate of human corneas was 25.5 micron/hr in GBR and significantly increased to 32.7 in BSS and 66.1 in NaCl. The epithelial intracellular potential of human cornea was constant at about 60 mV up to 5 hr of incubation in GBR and decreased continuously to 40 mV in BSS and NaCl. Endothelial cell potentials were stable for up to 3 hr of incubation in GBR or BSS and decreased from a control value of about 18 mV to 10 mV 2 hr after bathing in NaCl. Qualitatively similar data were obtained in the isolated rabbit cornea. The results demonstrate the advantage of GBR as an intraocular irrigating solution.

Animals

Evidence against phospholipid asymmetry in intracellular membranes from liver.

We have studied the distribution of phospholipids across the membrane of microsomal vesicles and Golgi-derived secretory vesicles from rat liver by the use of phospholipases. Model studies on single-bilayer phospholipid vesicles showed that phospholipase A2 (phosphatide 2-acyl-hydrolase, EC 3.1.1.4) cleaved at least 80% of the lipids on the outer surface of such vesicles without significant attack on the inner surface. In microsomal vesicles approximately 40% of the outer surface phospholipids were cleaved before the enzyme gained access to the interior of the vesicles. The same conclusion was reached for Golgi vesicles. By following the degradation of the three major phospholipids in intact microsomes and in extracted lipids we found that the same fraction of each of these phospholipids was exposed on the outer surface of the microsomal vesicles. Corresponding experiments with Golgi vesicles showed that distinctly different fractions of phosphatidylcholine and phosphatidylethanolamine were present on the surface of these vesicles. However, the difference was accounted for by enrichment of phosphatidylcholine in intravesicular particles rather than by asymmetry across the vesicle membrane. The results from specific hydrolysis of phosphatidylinositol confirmed an essentially symmetric distribution of this phospholipid across the microsomal and the Golgi vesicle membranes.

Animals

Intracellular membrane junctions during the exocytosis of insulin.

In the process of insulin release by exocytosis, a fusion between the membranes of secretory vesicles and the plasma membrane occurs which is followed by the rupture of these membranes and the formation of an exocytotic stoma. As revealed by freeze-fracturing, the membrane structure is altered in the region of membrane interaction. An accumulation of membrane-associated particles can be observed in membranes of secretory vesicles contacting the plasma membrane. At the exocytotic stoma the boundary between granule limiting membrane and plasma membrane is occasionally outlined by a ring-like aggregation of membrane-associated particles. The effects of hypertonic solutions on tissue structure indicate that the membrane junctions during exocytosis are permeable to ions and small molecules, forming a communication between the interior of the secretory vesicle and the extracellular space.

Animals