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Biomedical subjects

S Knutton

Publications and source records attributed to S Knutton.

83 records · Page 5Linked to original sources

The fluidity of normal and virus-transformed cell plasma membrane.

1. The phospholipid composition and cholesterol/phospholipid ratio of plasma membrane is the same in normal as in transformed BHK (baby-hamster kidney) cells; no significant difference in length or degree of unsaturation of the contributing acyl chains is apparent. 2. The turnover of acetate-labelled phosphatidylcholine species in the plasma membrane of normal and transformed BHK cells is the same. 3. Intramembranous particles of normal and transformed 3T3-cell plasma membrane are randomly distributed, whether at 4degreesC or at 37degreesC, in sparse or in dense cultures. There is no correlation between distribution of particles and the movement of concanavalin A receptor sites. 4. It is concluded that transformation of fibroblastic cells by oncogenic viruses does not lead to major changes in the lipid fluidity of the plasma membrane. 5. Details of the phospholipid composition of nuclei, mitochondria and endoplasmic reticulum in normal and transformed BHK cells have been deposited as Supplementary Publication SUP 50061 (5 pages) at the British Library Lending Division, Boston, Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies may be obtained on the terms given in Biochem. J. (1976) 153, 5.

Cell Membrane↗

Regular structures in unit membranes. III. Further observations on the particulate component of the suckling rat ileum endocytic membrane complex.

Further morphological observations on the particulate components decorating the lumenal surfaces of membranes of the endocytic complex of the epithelial cells of the suckling rat ileum are presented. The particles each measure approximately 7.5 nm across and give the appearance of the capital letter H in frontal view. They consist of the enzyme n-acetyl-beta-glucosaminidase (NAG). They are arranged in rows called "decorated strips" with the symmetrical lateral bars in register and spaced approximately 14.5 nm apart. Decorated strips lie side-by-side in the external (lumenal) surface of the membrane. They are parallel and sometimes spaced approximately 14.5 nm apart making an orthogonal lattice. The lateral spacing between the decorated strips under certain conditions is reduced and sometimes there is shear between the adjacent ones. Occasionally, shear is present within the decorated strips themselves, with slight displacement of the two sides of each H-shaped particle. A purified preparation of these membranes has been studied by electron microscopy using thin sectioning, negative stain, Markham translation and optical diffraction computer image reconstruction methods. The individual particles comprising the array can be seen in the membrane surface in profile view when dried in a pool of negative stain. They appear either triangular or diamond-shaped in such views. If triangular, they appear to consist of three domains at the corners of an equilateral triangle. One side of each triangular figure is parallel to the membrane surface but separated from it by a dense band of negative stain approximately 2 nm thick that runs along the surface of the membrane. Sometimes a fourth symmetrical domain is visible within this dense band, giving a diamond-shaped figure. This fourth domain connects the particle to the membrane. Thus, each H-shaped particle is a double structure, with each half in profile view appearing as a diamond figure of four symmetrical domains. Each H-shaped particle is believed to consist of either two or four molecules of NAG.

Animals↗

Regular structures in membranes: the lumenal plasma membrane of the cow urinary bladder.

The ultrastructure of the lumenal plasma membrane of the cow urianry bladder has been studied in thin sections of glutaraldehyde- and glutaraldehyde-H2O2-fixed specimens, by negative staining and freeze fracture. A regular hexagonal array of particles confined to polygonal plaques 0-1-0-4-mum in diameter and separated by 0-02-mum interplaque areas is revealed by all 3 techniques. Cross-sections through particulate areas fixed with glutarayldehyde-H2O2 display a tetralaminar structure consisting of the usual approximately 8-nm-thick trilamellar unit membrane structure, on the external dense leaflet of which is located an additional approximately 4-nm-thick stratum which is occasionally resolved into a row of regulrly spaced approximately 4-nm-diameter particles. Non-particulate areas feature only the approximately 8-nm-thick trilamellar structure. Tangential sections reveal an hexagonal array of particles with a unit cell of approximately 16 nm. Four membrane faces can be revealed by freeze fracture and etching of membranes of the cow urinary bladder; 2 complementary split inner membrane faces (A and B) revealed by the cleaving process and the lumenal and cytoplasmic membrane surfaces exposed by etching. Face B, which belongs to the external membrane leaflet and faces the cytoplasm, displays plaques of particles arranged in a hexagonal lattice with a unit cell of approximately 16 nm. Face A, which belongs to the cytoplasmic membrane leaflet and faces the lumen, displays a complementary array of hexagonally packed pits. The hexagonally arranged particles also protrude into the lumenal membrane surface where they can occasionally be resolved into 6 approximately 5-nm-diameter subunits; the cytoplasmic surface appears smooth. Six approximately 5-nm-diameter subunits are also revealed in negatively stained preparations. The data are consistent with a model for the membrane in which the particles forming the hexagonal structure protrude above the lumenal membrane surface and also bridge most of the thickness of the membrane.

Animals↗

Role of microvilli in surface changes of synchronized P815Y mastocytoma cells.

The surface morphology of synchronized P815Y mastocytoma cells has been examined by scanning electron microscopy. Early G1 cells are comparatively smooth or light villated, whereas at later stages the surface becomes progressively more villated. In G1 cell most microvilli have a uniform diameter, whereas in S and G2 cells, many microvilli show branching and often originate from much larger surface protuberances. Small "blebs" are seen on the surface of many cells but these structures do not appear to be a characteristic feature of cells at any one stage of the cycle. The presence of microvilli increases the total surface of the cell to such an extent that the ratio of volume to surface area remains constant throughout the cell cycle. The mechanism of cytokinesis is thus a physical one, involving the unfolding of previously accumulated microvilli.

Cell Division↗

The perturbation of the human erythrocyte membrane by phospholipase C.

A study has been made of freeze-fractured preparations of erythrocyte ghosts modified by phospholipase C (Clostridium welchii). Such membranes show a decrease in surface area of up to about 47% and lipid droplets appear on their external surface but there is no loss of protein. Freeze-fracture of maximally hydrolysed membranes exposes only very small areas of A faces and these appear particle-free. Most of the membranes are simply cross-fractured. At lower levels of hydrolysis there is more extensive exposure of A fracture faces but the particle density is less than in control preparations. If such exposed faces were representative of the whole membrane then the particle density would have been expected to increase. It is suggested either that areas of membrane with increased particle density do not fracture or that the particles revealed by freeze-fracture involve phospholipid as well as protein and are not revealed in the absence of phospholipid.

Blood Proteins↗

Regular structures in membranes. I. Membranes in the endocytic complex of ileal epithelial cells.

An "apical endocytic complex" in the ileal lining cells of suckling rats is described. The complex consists of a continuous network of membrane-limited tubules which originate as invaginations of the apical plasma membrane at the base of the microvilli, some associated vesicles, and a giant vacuole. The lumenal surface of this tubular network of membranes and associated vesicles is covered with a regular repeating particulate structure. The repeating unit is an approximately 7.5-nm diameter particle which has a distinct subunit structure composed of possibly nine smaller particles each approximately 3 nm in diameter. The approximately 7.5-nm diameter particles are joined together with a center-to-center separation of approximately 15 nm to form long rows. These linear aggregates, when arranged laterally, give rise to several square and oblique two-dimensional lattice arrangements of the particles which cover the surface of the membrane. Whether a square or oblique lattice is generated depends on the center-to-center separation of the rows and on the relative displacement of the particles in adjacent rows. Four membrane faces are revealed by fracturing frozen membranes of the apical tubules and vesicles: two complementary inner membrane faces exposed by the fracturing process and the lumenal and cytoplasmic membrane surfaces revealed by etching. The outer membrane face reveals a distinct array of membrane particles. This array also sometimes can be seen on the outer (B) fracture face and is sometimes faintly visible on the inner (A) fracture face. Combined data from sectioned, negatively stained, and freeze-etched preparations indicate that this regular particulate structure is a specialization that is primarily localized in the outer half of the membrane mainly in the outer leaflet.

Animals↗

The molecular organization of lipids in the membrane of Escherichia coli: phase transitions.

X-ray diffraction techniques have been used to investigate the physical state of the lipids in the membrane of an unsaturated fatty acid auxotroph of Escherichia coli. Thermotropic phase transitions have been detected in membranes prepared from cells grown on various fatty acid supplements. Below the transition temperature, the X-ray diffraction pattern features a sharp ring at 420 pm (4.2 A) due to the close hexagonal packing of the apolar groups of the lipids; above the transition temperature the lipids are in a less organized liquid-crystalline state that gives rise to a diffuse band centered at a Bragg spacing of 460 pm (4.6 A). This transition occurs at or below the temperature at which the cells were grown. Disparities between this transition temperature and the temperature of discontinuities in the Arrhenius plots for proline transport and succinic dehydrogenase activity lead us to conclude that the distribution of lipids within the membrane is heterogeneous.

Cell Membrane↗