The structure and associations of the double S layer on the cell wall of Aquaspirillum sinuosum

1990 ◽  
Vol 36 (5) ◽  
pp. 327-335 ◽  
Author(s):  
Stephen H. Smith ◽  
Robert G. E. Murray
Keyword(s):  
Cell Wall ◽  
Outer Membrane ◽  
Outer Layer ◽  
Peptide Mapping ◽  
Periodic Acid ◽  
Minor Component ◽  
Membrane Resistance ◽  
Surface Layers ◽  
Periodic Acid Schiff ◽  
Bacterial Surface

Aquaspirillum sinuosum cell walls bear two paracrystalline, proteinaceous surface layers (S layers). Each shows a different symmetry: the inner layer is closely apposed to the outer membrane and is a tetragonal array (90° axes; 5-nm units; repeat frequency 8 nm); the outer layer is a hexagonal array on the external surface (14-nm units; repeat frequency 18 nm) and, although the units have a six-pointed stellate form, the linkage between units is not resolved. The outer layer consists of a major 130-kDa protein and a 180-kDa minor component; these co-extract, co-assemble, and are inseparable by hydroxylapatite chromatography or by recrystallization. The solubilizing effects of reagents suggest stabilization by hydrogen bonding and Ca2+. The two outer layer proteins are serologically related and show partial identity by peptide mapping. Periodic acid – Schiff staining of the 180-kDa band suggests that this may be a glycosylated form of the 130-kDa component. The inner layer components form a doublet of 75- and 80-kDa polypeptides with extreme resistance to extraction. Close apposition to the outer membrane, resistance to chaotropes, aqueous insolubility, and behaviour in charge-shift electrophoresis suggest hydrophobic interaction between subunits and an integral association with the outer membrane. Key words: bacterial surface, cell wall, surface layers, cell-wall proteins, cell-wall assembly.

Prefertilization ovule development in Capsella: the dyad, tetrad, developing megaspore, and two-nucleate gametophyte

1986 ◽  
Vol 64 (4) ◽  
pp. 875-884 ◽  
Author(s):  
Patricia Schulz ◽  
William A. Jensen
Keyword(s):  
Cell Wall ◽  
Electron Microscope ◽  
Acid Phosphatase ◽  
De Novo ◽  
Periodic Acid ◽  
Aniline Blue ◽  
Ovule Development ◽  
Periodic Acid Schiff ◽  
Fluorescent Material ◽  
Autophagic Vacuoles

Ovules of Capsella bursa-pastoris at the dyad and tetrad stages of meiosis and at the megaspore and two-nucleate stages of the gametophyte were studied with the electron microscope. The cells of the dyad and tetrad are separated by aniline blue fluorescent cross walls and receive all types of organelles and autophagic vacuoles that were present in the meiocyte. Autophagic vacuoles enclose ribosomes and organelles and show reaction product for acid phosphatase. Autophagic vacuoles and some plastids are absorbed into the enlarging vacuoles of the growing megaspore. Other plastids appear to survive meiosis and there is no evidence for their de novo origin. Some mitochondria appear to degenerate in the enlarging megaspore but others look healthy and there is no evidence for the de novo origin of mitochondria. The nucleolus of the developing megaspore becomes very large and the cytoplasm is extremely dense with ribosomes. The cell wall is thickened by an electron-translucent, periodic acid – Schiff negative, aniline blue fluorescent material and contains plasmodesmata that link the megaspore with the nucellus. The plasmalemma of the growing megaspore produces microvilluslike extensions into this wall that disappear with the formation of the two-nucleate gametophyte. Plasmodesmata disappear from the cell wall at the four-nucleate stage.

scholarly journals The lanthanide-enhanced affinity chromatography of clostridial collagenase

1985 ◽  
Vol 225 (2) ◽  
pp. 553-556 ◽  
Author(s):  
C H Evans
Keyword(s):  
Periodic Acid ◽  
Sodium Dodecyl ◽  
Minor Component ◽  
Electrophoretic Analysis ◽  
Lanthanide Ions ◽  
Affinity Column ◽  
Major Protein ◽  
Periodic Acid Schiff ◽  
Highly Active ◽  
A Minor

Clostridiopeptidase A (EC 3.4.24.3) did not bind to a collagen affinity column in the absence of Ca2+, but did so in the presence of lanthanide ions (Ln3+). The sequestered enzyme could be eluted with EGTA. For the four Ln3+ ions tested, the order of efficiency in promoting enzyme binding, Sm3+ greater than Lu3+ greater than Er3+ much greater than La3+, reflected their relative abilities to inhibit clostridiopeptidase A. By using Sm3+ as an adjunct, it proved possible to separate a highly active preparation of collagenase from crude clostridial collagenase. Sodium dodecyl sulphate/polyacrylamide-gel-electrophoretic analysis of the preparation revealed a major protein of Mr 95000 and a minor component of Mr 82000. As both were stained by periodic acid/Schiff reagent, they were probably glycoproteins.

Cell surface characteristics of Mortierella species and their interaction with a mycoparasite

1984 ◽  
Vol 30 (3) ◽  
pp. 290-298 ◽  
Author(s):  
M. S. Manocha
Keyword(s):  
Cell Wall ◽  
Cell Surface ◽  
Periodic Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Surface Characteristics ◽  
Protein Patterns ◽  
Periodic Acid Schiff ◽  
The Difference ◽  
Chitin And Chitosan

Cell surface characteristics of three Mortierella species differing in their response to a mycoparasite, Piptocephalis virginiana, were examined. Their cell wall composition was typical of mucoraceous fungi with chitin and chitosan as major polysaccharides. Electron microscopy revealed that the mycoparasite penetrated and formed haustoria in the hyphae of susceptible hosts, M. pusilla and M. isabellina. The failure of the parasite to establish contact and penetrate a hypha of the nonhost, M. candelabrum, was not due to cell wall thickness, rigidity, or chitin contents. Markedly different protein patterns obtained from crude alkali extracts of host and nonhost cell walls by sodium dodecyl sulfate – polyacrylamide gel electrophoresis might explain the difference in host and nonhost response to the mycoparasite. Whereas most of the bands differed only in intensity after staining with either Coomassie blue or periodic acid – Schiff reagent, there were two distinct bands of glycoproteins (76 000 and 74 000) observed in the host species which were absent in the nonhost species.

scholarly journals Structural and kinetic properties of a novel purple acid phosphatase from phosphate-starved tomato (Lycopersicon esculentum) cell cultures

2004 ◽  
Vol 377 (2) ◽  
pp. 419-428 ◽  
Author(s):  
Gale G. BOZZO ◽  
Kashchandra G. RAGHOTHAMA ◽  
William C. PLAXTON
Keyword(s):  
Acid Phosphatase ◽  
Lycopersicon Esculentum ◽  
Peptide Mapping ◽  
Periodic Acid ◽  
Purple Acid Phosphatase ◽  
Kinetic Properties ◽  
Suspension Cells ◽  
Periodic Acid Schiff ◽  
Α Subunit ◽  
Α And Β Subunits

An intracellular acid phosphatase (IAP) from Pi-starved (−Pi) tomato (Lycopersicon esculentum) suspension cells has been purified to homogeneity. IAP is a purple acid phosphatase (PAP), as the purified protein was violet in colour (λmax=546 nm) and was insensitive to l-tartrate. PAGE, periodic acid–Schiff staining and peptide mapping demonstrated that the enzyme exists as a 142 kDa heterodimer composed of an equivalent ratio of glycosylated and structurally dissimilar 63 (α-subunit) and 57 kDa (β-subunit) polypeptides. However, the nine N-terminal amino acids of the α- and β-subunits were identical, exhibiting similarity to the deduced N-terminal portions of several putative plant PAPs. Quantification of immunoblots probed with rabbit anti-(tomato acid phosphatase) immune serum revealed that the 4-fold increase in IAP activity due to Pi-deprivation was correlated with similar increases in the amount of antigenic IAP α- and β-subunits. IAP displayed optimal activity at pH 5.1, was activated 150% by 10 mM Mg2+, but was potently inhibited by Zn2+, Cu2+, Fe3+, molybdate, vanadate, fluoride and Pi. Although IAP demonstrated broad substrate selectivity, its specificity constant (Vmax/Km) with phosphoenolpyruvate was >250% greater than that obtained with any other substrate. IAP exhibited significant peroxidase activity, which was optimal at pH 9.0 and insensitive to Mg2+ or molybdate. This IAP is proposed to scavenge Pi from intracellular phosphate esters in −Pi tomato. A possible secondary IAP role in the metabolism of reactive oxygen species is discussed. IAP properties are compared with those of two extracellular PAP isoenzymes that are secreted into the medium of −Pi tomato cells [Bozzo, Raghothama and Plaxton (2002) Eur. J. Biochem. 269, 6278–6286].

Cell wall proteins of Candida albicans

1983 ◽  
Vol 29 (10) ◽  
pp. 1438-1444 ◽  
Author(s):  
W. LaJean Chaffin ◽  
Douglas M. Stocco
Keyword(s):  
Cell Wall ◽  
Candida Albicans ◽  
Periodic Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Periodic Acid Schiff ◽  
Temperature Growth ◽  
Growth State ◽  
Yeast Cultures ◽  
Schiff Reagent ◽  
Germ Tubes

Proteins were solubilized from cell wall fractions of Candida albicans and separated by polyacrylamide gel electrophoresis. Cell walls were isolated from 25 and 37 °C growing and stationary phase yeast cultures and from germ tubes. The 42 protein bands detected by dye binding were observed in all wall extracts, regardless of the temperature, growth state, or morphology of the culture. The carbohydrate content of most bands was below the detectable limit of the periodic acid Schiff reagent. The protein complement revealed by autoradiography of radiolabeled proteins was half that detected by staining. Two bands showed greater intensity from cultures grown at 37 °C. The radio-labeled pattern was similar with both [35S]methionine- and [14C]leucine-labeled proteins and either pulse- or continuous-labeled proteins.

The surface structure of Leptotrichia buccalis

1994 ◽  
Vol 40 (2) ◽  
pp. 90-98 ◽  
Author(s):  
S. H. Smith ◽  
R. G. E. Murray ◽  
M. Hall
Keyword(s):  
Cell Wall ◽  
Surface Structure ◽  
Outer Membrane ◽  
Room Temperature ◽  
Minor Component ◽  
Human Group ◽  
Ridge Structure ◽  
Sds Page ◽  
In Series ◽  
Group A

Leptotrichia buccalis shows a mosaic of surface structure on its outer membrane consisting of curved ridges 35 nm high and 22 nm apart, and erect on that surface. Fimbriae (common pili) are not present and nor is an S layer. The flap-like ridges consist of strings of macromolecules radiating from the cell surface. This ridge structure is not soluble in any of the usual chaotropes and can only be released when the outer membrane has been damaged or dispersed by extracting envelope preparations with 0.5% SDS at room temperature. The ridge is then found to be attached firmly to the peptidoglycan sacculus, which may be the point of origin of the structure. When so prepared the macromolecules forming the ridge can be removed from the sacculus by treatment with 6 M guanidine HCl, and SDS-PAGE analysis of the extract reveals a 210-kDa polypeptide as a major component and a 15-kDa minor component. The latter is probably a peptidoglycan-associated protein and much of it remains with the sacculus. Each string forming the ridge is of a volume consistent with being made of three elongated 210-kDa molecules, which are united in series by strong hydrophobic association and laterally with neighboring strings by slightly weaker forces. We confirm that L. buccalis causes haemagglutination and the bacteria are known to attach to various tissue cells. Human group A red blood corpuscles remove both of the proteins from solution, which supports the hypothesis that the ridges are adhesin structures. It is likely but not proven that the 210-kDa molecule is the adhesin.Key words: Leptotrichia buccalis, cell wall, S layer, protein array, adhesin.

scholarly journals Surface organization and composition of Euglena. II. Flagellar mastigonemes

1978 ◽  
Vol 77 (3) ◽  
pp. 805-826 ◽  
Author(s):  
GB Bouck ◽  
A Rogalski ◽  
A Valaitis
Keyword(s):  
Unit Length ◽  
Periodic Acid ◽  
Minor Component ◽  
Positive Staining ◽  
Periodic Acid Schiff ◽  
Phenol Extraction ◽  
Low Concentrations ◽  
Flagellar Membrane ◽  
Pentose Sugar ◽  
Layer Chromatography

The surface of the Euglena flagellum is coated with about 30,000 fine filaments of two distinct types. The longer of these nontubular mastigonemes (about 3 micron) appear to be attached to the paraflagellar rod whereas the shorter nontubular mastigonemes (about 1.5 micron) are the centrifugally arranged portions of a larger complex, which consists of an attached unit parallel to and outside of the flagellar membrane. Units are arranged laternally in near registration and longitudinally overlap by one-half of a unit length. Rows of mastigoneme units are firmly attached to the axoneme microtubules or to the paraflagellar rod as evidenced by their persistence after removal of the flagellar membrane with neutral detergents. SDS-acrylamide gels of whole flagella revealed about 30 polypeptides, of which two gave strong positive staining with the periodic acid-Schiff (PAS) procedure. At least one of these two bands (glycoproteins) has been equated with the surface mastigonemes by parallel analysis of isolated and purified mastigonemes, particularly after phenol extraction. The faster moving glycoprotein has been selectively removed from whole flagella and from the mastigoneme fraction with low concentrations of neutral detergents at neutral or high pH. The larger glycoprotein was found to be polydisperse when electrophoresed through 1% agarose/SDS gels. Thin-layer chromatography of hydrolysates of whole flagella or of isolated mastigonemes has indicated that the major carbohydrate moiety is the pentose sugar, xylose, with possibly a small amount of glucose and an unknown minor component.

Ultrastructure of Enterococcus faecalis biofilms

Biofilms ◽  
2004 ◽  
Vol 1 (2) ◽  
pp. 131-137 ◽  
Author(s):  
S. L. Erlandsen ◽  
C. J. Kristich ◽  
G. M. Dunny
Keyword(s):  
Cell Wall ◽  
Enterococcus Faecalis ◽  
Biofilm Formation ◽  
Single Cells ◽  
Periodic Acid ◽  
Bacterial Cells ◽  
Dialysis Tubing ◽  
Periodic Acid Schiff ◽  
Kidney Dialysis ◽  
Polystyrene Surface

Enterococcus faecalis is known to produce biofilms on biomaterials, but the manner in which this occurs is unknown. Herein we report that adhesion of E. faecalis in biofilms appeared to be mediated by cell wall surface projections attaching cells to the substratum. Biofilm formation was observed on the polystyrene surface of 96-well plates and also on the surface of cellulose kidney dialysis tubing used as a model for biofilm formation on catheters. Qualitative differences involved the packing of E. faecalis cells in biofilms, with greater intercellular spacing detected in the 96-well plate, whereas bacteria were tightly packed on the surface of cellulose catheters. Distribution of adherent bacterial cells accumulating on the two surfaces revealed obvious differences, with most of the bacteria attaching to the polystyrene surface as single cells or diplococci separated from neighboring organisms by intervals of uncolonized surface. In contrast, enterococci on the cellulose surface were found as multi-layer cellular aggregates or microcolonies, even when much of the total surface was free from attached bacteria. Microcolonies stained intensely for neutral hexose sugars using the periodic acid–Schiff (PAS) stain. Surface projections, presumably exopolysaccharide, anchored bacteria to the substratum and appeared to elevate the cells above the surface. These slender surface projections could be seen over the entire enterococcal cell wall, with the exception of areas adjacent to septal regions where new cell wall formation was occurring. Rod-like interconnections were also observed between adjacent diplococci. These results suggested that biofilm formation varies on different substrates and that enterococcal surface projections may be involved in E. faecalis colonization and adhesion within biofilms.

Ultrastructure of the Parotid Gland of the Nine-Banded Armadillo

1977 ◽  
Vol 35 ◽  
pp. 640-641
Author(s):  
J. R. Ruby
Keyword(s):  
Endoplasmic Reticulum ◽  
Parotid Gland ◽  
Periodic Acid ◽  
Acinar Cells ◽  
Duct System ◽  
Parotid Glands ◽  
Dasypus Novemcinctus ◽  
Anterior Portion ◽  
Periodic Acid Schiff ◽  
Thin Sections

Parotid glands were obtained from five adult (four male and one female) armadillos (Dasypus novemcinctus) which were perfusion-fixed. The glands were located in a position similar to that of most mammals. They extended interiorly to the anterior portion of the submandibular gland.In the light microscope, it was noted that the acini were relatively small and stained strongly positive with the periodic acid-Schiff (PAS) and alcian blue techniques, confirming the earlier results of Shackleford (1). Based on these qualities and other structural criteria, these cells have been classified as seromucous (2). The duct system was well developed. There were numerous intercalated ducts and intralobular striated ducts. The striated duct cells contained large amounts of PAS-positive substance.Thin sections revealed that the acinar cells were pyramidal in shape and contained a basally placed, slightly flattened nucleus (Fig. 1). The rough endoplasmic reticulum was also at the base of the cell.

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