scholarly journals Antibodies against Sulphoquinovosyl-diacyl glycerol and their Reactions with Chloroplasts

1970 ◽  
Vol 25 (4) ◽  
pp. 412-419 ◽  
Author(s):  
Alfons Radunz ◽  
Richard Berzborn
Keyword(s):  
Bovine Serum Albumin ◽  
Ultrasonic Treatment ◽  
Sucrose Gradient ◽  
Osmotic Shock ◽  
Antirrhinum Majus ◽  
Coombs Test ◽  
Phosphatidyl Glycerol ◽  
Lamellar System ◽  
Intravenous Injections ◽  
Fucus Serratus

Using immunological techniques we tried to determine the localization and orientation of the sulpholipid of diloroplasts in thylakoids. During immunization of rabbits with lamellar systems of chloroplasts of Antirrhinum majus antisera were obtained which precipitated sulpholipid emulsions. Antibodies against the sulpholipid were not detected until a second series of intravenous injections.By immunizing purified sulpholipid of Fucus serratus, adsorbed to methylated bovine serum albumin, sulpholipid antisera were acquired. These antisera and the antisera against lamellar systems of Antirrhinum majus reacted with preparations of sulpholipid from widely different plant species. The sulpholipid antisera did not react with emulsions of the mono- or digalactosyl-diacyl glycerol, of lecithin or the anionic phosphatide phosphatidyl glycerol. Lamellar systems isolated without osmotic shock on a sucrose gradient were not agglutinated by sulpholipid antisera directly, but an indirect agglutination could be achieved in a Coombs-test or in a mixed antigen agglutination after Uhlenbruck. Broken chloroplasts, however, were agglutinated directly by sulpholipid antisera, as well lamellar systems which had been disrupted in a French-press or by ultrasonic treatment.From agglutination and agglutination inhibition experiments with lamellar systems prepared by different methods it is concluded that the determinant groups of the sulpholipid are accessible for antibodies in the lamellar system of chloroplasts. But probably the sulpholipid molecules are located inside the thylakoids.

Binding of Antibodies onto the Thylakoid Membrane I. Maximal Antibody Binding and Adsorption of Antibodies to Lipids

1975 ◽  
Vol 30 (7-8) ◽  
pp. 484-488 ◽  
Author(s):  
Alfons Radunz
Keyword(s):  
Surface Structure ◽  
Thylakoid Membrane ◽  
Serum Proteins ◽  
Antibody Binding ◽  
Molecular Surface ◽  
Antirrhinum Majus ◽  
Saturation Curve ◽  
Phosphatidyl Glycerol ◽  
Lamellar System ◽  
Monogalactosyl Diglyceride

Abstract The binding of antibodies onto the lamellar system of Antirrhinum majus was determined in dependence on the serum addition. The unspecific adsorption of serum proteins was taken into account or eliminated. The binding of antibodies as a function of the amount of serum added is seen from a saturation curve. From an antiserum obtained by hyperimmunization with stroma-freed chloroplasts, the chloroplasts bind maximally 1 gram antibodies per gram stroma-freed chloro­ plasts. From an antiserum to the proteins of the thylakoid membrane prepared in the same way an equal amount of antibodies is adsorbed. It is assumed that with this amount the surface of the lamellar system accessible to antibodies is completely covered by antibodies. For an antiserum to monogalactosyl diglyceride a maximal antibody binding of 0.16 g, for sulphoquinovosyl diglyceride 0.12 g and for phosphatidyl glycerol 0.13 g of antibodies per gram stroma-freed chloroplasts are obtained. The significance of these results with respect to the molecular surface structure of the thylakoid membrane is discussed.

Phosphatidylglycerin-Antiserum und seine Reaktionen mit Chloroplasten / Phosphatidyl glycerol Antiserum and its Reactions with Chloroplasts

1971 ◽  
Vol 26 (9) ◽  
pp. 916-919 ◽  
Author(s):  
Alfons Radunz
Keyword(s):  
Chloride Solution ◽  
Sucrose Solution ◽  
Thylakoid Membranes ◽  
Tris Buffer ◽  
Coombs Test ◽  
Phosphatidyl Glycerol ◽  
Protein Layer ◽  
Lamellar System ◽  
Accessible Surface ◽  
Isolated Chloroplasts

By immunisation of rabbits with stroma-freed chloroplasts and other preparations of the lamellar system, antibodies to phosphatidyl glycerol were formed. By injection of phosphatidyl glycerol, bound to methylated bovine serum albumin, the rabbits also reacted with the formation of antibodies to this phosphatide. The antiserum to phosphatidyl glycerol yielded no reactions with phosphatidyl choline and the glycolipids sulphoquinovosyl diglyceride and mono- and digalactosyl diglyceride, which are present in chloroplasts.Stroma-freed chloroplasts from Antirrhinum majus and Nicotiana tabacum were not agglutinated by antisera to phosphatidyl glycerol. By means of the Coombs test, the “mixed-antigen-agglutination” and sateration experiments it was possible to demonstrate that antibodies to phosphatidyl glycerol are specifically adsorbed onto stroma-freed chloroplasts. After partial protein decomposition by proteases, stroma-freed chloroplasts were agglutinated. Fragments of the thylakoid membranes (ultrasonic supernatant) were precipitated only after partial protein decomposition and functioned in the Coombs test just like stroma-freed chloroplasts. However, fragments of the lamellar system (ultrasonic sediment) were directly agglutinated. Isolated chloroplasts in tris buffer-sucrose solution and in tris buffer-sodium chloride solution were equally agglutinated by antiserum to phosphatidyl glycerol.From the experiments it is concluded that the antigen determinants of the phosphatidyl glycerol are located on the antibody-accessible surface of the thylakoids. The agglutination of stroma-freed chloroplasts was, however, sterically hindered by membrane proteins. In those cases in which chloroplast preparations were directly agglutinated, it is assumed that either thylakoids were disrupted or that they were swollen, resulting in a change in the protein layer of the membranes in such a way that no steric hindrance occurs.

Membranes in lupin root nodules. II. Preparation and properties of peribacteroid membranes and bacteroid envelope inner membranes from developing lupin nodules

1978 ◽  
Vol 30 (1) ◽  
pp. 151-174
Author(s):  
J.G. Robertson ◽  
M.P. Warburton ◽  
P. Lyttleton ◽  
A.M. Fordyce ◽  
S. Bullivant
Keyword(s):  
Sucrose Gradient ◽  
Phosphotungstic Acid ◽  
Nadh Oxidase ◽  
Osmotic Shock ◽  
Root Nodules ◽  
Freeze Fracture ◽  
Electrophoretic Analysis ◽  
Thin Sections ◽  
Peribacteroid Space ◽  
Gradient Fractionation

Peribacteroid membranes and bacteroid envelope inner membranes have been isolated from developing lupin nodules. Isolation of the peribacteroid membranes was achieved by first preparing membrane-enclosed bacteroids free from other plant organelles or membranes. The peribacteroid membranes were then released by osmotic shock and purified by centrifugation to equilibrium on sucrose gradients. The bacteroids were broken in a pressure cell and the bacteroid envelope inner membranes were isolated using sucrose gradient fractionation of the bacteroid total envelope preparation. The density of the peribacteroid membranes decreased during the period of development of N2-fixation in lupin nodules from 1.148 g/ml for nodules from 12-day plants to 1.137 g/ml for nodules from 18-day plants. The density of the bacteroid envelope inner membranes from nodules from 18-day plants was 1–153 g/ml. The identity and homogeneity of the isolated membranes was established, by comparison with membranes in intact nodules, using phosphotungstic acid and silver staining of thin sections and particle densitites on faces of freeze-fracture replicas of the membranes. Analyses for NADH oxidase and succinate dehydrogenase, spectral analyses and gel-electrophoretic analysis of proteins were also used to characterize the membrane and soluble protein fractions from the nodules. The ratio of lipid to protein was 6.1 for the peribacteroid membranes and 2.5 for the bacteroid envelope inner membranes. Leghaemoglobin was localized in the plant cytoplasm in lupin nodules and not in the peribacteroid space.

Konzentration einiger Lipide in den Chloroplasten von Zea mays und Antirrhinum majus / Concentration of Some Lipids in the Chloroplasts of Zea mays and Antirrhinum ma jus

1971 ◽  
Vol 26 (11) ◽  
pp. 1180-1187 ◽  
Author(s):  
Friederike Koenig
Keyword(s):  
Zea Mays ◽  
Aqueous Medium ◽  
Vitamin K ◽  
Lipid Content ◽  
Dry Weight ◽  
Antirrhinum Majus ◽  
Lamellar System ◽  
Total Leaf

Stroma-containing chloroplasts from Zea mays and Antirrhinum majus were isolated in aqueous medium. The average dry weight of chloroplasts from Zea mays is 27·10-12 g, that of Antirrhinum majus 30·10-12 g. Water freed chloroplasts consist up to 49 or 45 percent respectively of lamellar system. The lipid content of the lamellar system of Zea mays is 49 percent, that of Antirrhinum majus 45 percent. A chloroplast of Zea mays contains on the average 920·106 chlorophyll molecules, 220·106 carotenoid molecules, 2000·106 molecules of galactolipids, 190·106 molecules of sulpholipid, 260·106 phosphatide molecules and 64·106 molecules of lipophilic quinones. In addition to phosphatidylglycerol also phosphatidylinositol and phosphatidylcholine were found. It is very probable that besides vitamin K1 the homologeous compound lacking one methylgroup is present in the chloroplasts. In contrast to the literature only 62 percent of the total leaf galactolipids are found in the chloroplasts.

Lokalisierung des Monogalaktosyldiglycerids in Thylakoidmembranen mit serologischen Methoden / Serological Localisation of Monogalactosyl Diglyceride in Thylakoid Membranes

1972 ◽  
Vol 27 (7) ◽  
pp. 822-826 ◽  
Author(s):  
Alfons Radunz
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Thylakoid Membrane ◽  
Bovine Serum ◽  
Thylakoid Membranes ◽  
Specific Antiserum ◽  
Lamellar System ◽  
Chloroplast Membrane ◽  
Monogalactosyl Diglyceride

Immunoglobulins to monogalactosyl diglyceride have been found in antisera to different chloroplast preparations. A specific antiserum to this lipid has been obtained by injection of monogalactosyl diglyceride which was adsorbed onto methylated bovine serum albumin. Inhibition tests with mono- and oligosaccharides, as well as with mono- and digalactosyl glycerol, showed that the immunoglobulin was directed to the β-galactosyl-(1 → 1) -glycerol configuration. Digalactosyl diglyceride and sulphoquinovosyl diglyceride did not react with the antiserum. The immunoglobulin is of the Ig M-type.The antiserum to monogalactosyl diglyceride agglutinates stroma-freed chloroplasts. Fragments of the thylakoid membrane, obtained by ultrasonication and fractioning centrifugation, thylakoids and thylakoid stacks are all precipitated. In a preparation of stroma-containing chloroplasts agglutination takes place but after rupture of the chloroplast membrane and loss of the stroma. The ultrasonicated lamellar system binds a larger amount of the monogalactosyl diglyceride immunoglobulins than does stroma-freed chloroplasts. It is concluded from these experiments that some of the chloroplasts monogalactosyl diglyceride is located at the surface of the thylakoids.

Studies On The Structure And Cellular Location Of Various Ribosome And Ribosomal Rna Species In The Green Alga Chlamydomonas Reinhardi

1971 ◽  
Vol 8 (1) ◽  
pp. 153-183 ◽  
Author(s):  
D. P. BOURQUE ◽  
J. E. BOYNTON ◽  
N. W. GILLHAM
Keyword(s):  
Ribosomal Rna ◽  
Gel Electrophoresis ◽  
Sucrose Gradient ◽  
Chlorophyll Synthesis ◽  
Sedimentation Velocity ◽  
Wild Type ◽  
Progressive Reduction ◽  
Lamellar System ◽  
Subunit Dissociation ◽  
Type C

Under ionic conditions effecting little or no subunit dissociation, Chlamydomonas reinhardi contains 2 major classes of ribosomes with generic sedimentation velocities of 83 and 70s and 3 minor classes with sedimentation velocities of 66, 54, and 41s. Ribosomal RNAs with sedimentation velocities of 25, 23, 18, 16 and 5s have been identified. The 70-s ribosomes are in the chloroplast and contain 23-, 16- and 5-s ribosomal RNA whereas the 83-s ribosomes are in the cytoplasm and contain 25-, 18- and 5-s ribosomal RNA. Numbers of chloroplast ribosome particles counted in electron micrographs of wild type C. reinhardi and the ac-20 and y mutants have been compared with relative amounts of 70-s ribosomes determined by sucrose gradient sedimentation and amounts of 23-, 16- and 5-s ribosomal RNA determined by gel electrophoresis. In response to reduced concentrations of magnesium the 70-s ribosomes of wild type are susceptible to a progressive reduction in sedimentation velocity whereas the 66-s ribosomes of the mutant ac-20 are not. Chlorophyll synthesis and the formation of the chloroplast lamellar system do not appear to be correlated with the relative amounts of chloroplast ribosomes.

Passage of bovine serum albumin from the mother to rabbit blastocysts

Development ◽  
1973 ◽  
Vol 30 (2) ◽  
pp. 459-469
Author(s):  
Floy L. Crutchfield ◽  
Abraham C. Kulangara
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Double Diffusion ◽  
Early Gestation ◽  
Uterine Lumen ◽  
Intravenous Injections ◽  
Uterine Fluid ◽  
Kinetics Of ◽  
Permeability Rate

Dutch belted rabbits were given single intravenous injections of 100 or 200 mg/kg doses of bovine serum albumin (BSA). BSA in serum and uterine fluid at various times after injection was estimated by a quantitative radial immunodiffusion test, which could measure a minimum of 40 ng. The presence of BSA in uterine fluid was confirmed by immunoelectrophoresis and double diffusion in agar. BSA passes readily into uterine fluid of non-pregnant rabbits, reaching a peak at 12 h after injection, when its concentration is 7–15% of that in serum. About 72 h seems to be required for equilibration of BSA between serum and uterine fluid, at which time the concentration in the former is about 5 times that in the latter. The kinetics of the process is discussed. Compared to the above, passage of BSA into uterine fluid of pregnant rabbits (5–7 days post coitum) is restricted in the following ways. Significant amounts of BSA appear in the fluid only after a maternal dose of 200 mg/kg. BSA in uterine fluid reaches a peak at 24 h after injection, when it is only 4·5% of the serum level. The permeability rate seems to decrease with early gestation. Approximate rates of entry of BSA into uterine lumen of non-pregnant and pregnant rabbits are 0·4 and 0·25 μg/h. BSA seems to be treated like rabbit albumin in its passage across the uterine epithelium. There is no evidence of selection between these proteins.

scholarly journals HISTOLOGICAL AND SEROLOGICAL SEQUENCES IN EXPERIMENTAL HYPERSENSITIVITY

1947 ◽  
Vol 85 (6) ◽  
pp. 571-590 ◽  
Author(s):  
Clinton Van Zandt Hawn ◽  
Charles A. Janeway
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Gamma Globulin ◽  
Horse Serum ◽  
Protein Solutions ◽  
Pathological Lesions ◽  
Intravenous Injections ◽  
Foreign Proteins ◽  
Antigen Antibody

1. Groups of normal rabbits were given, single intravenous injections of foreign proteins in doses of 1 gm. per kilo, bled at regular intervals for serologic studies, and sacrificed after varying lengths of time for pathological studies. The protein solutions used were of crystallized bovine serum albumin, bovine serum gamma globulin, and bovine serum. The experiments were planned, first, to correlate the sequence of pathological and immunological changes, and second, to compare the responses to two chemically and immunologically distinct plasma protein fractions and to the whole serum of the same species. 2. (a) The principal pathological lesions in rabbits given bovine serum were similar to those which have been previously observed following, the injection of horse serum and were characterized by widely dispersed but segmental acute inflammatory lesions of the arteries. These lesions were at their height 2 weeks after injection and showed marked repair at 4 weeks. (b) Crystallized bovine serum albumin produced lesions almost exclusively confined to the arteries which were at their height at 2 weeks, were healing at 3, and healed by 4 weeks. The lesions were less numerous and less intense than in animals given whole serum and were only found in some of the animals. (c) Bovine serum gamma globulin elicited quite different histologic sequences. The most striking lesions involved the glomeruli of the kidneys, and to a lesser degree, the heart. Lesions in the liver and joints were present but less conspicuous, and arterial lesions were rare and slight in degree. The lesions not only differed from those in rabbits given albumin in distribution but in timing, since they were most widespread and acute at 1 week and were healing at 2 weeks after injection. Moreover, lesions were observed in almost every animal. 3. Results of immunological studies were consistent with the interpretation that the pathological lesions were due to an antigen-antibody reaction in the tissues, as shown by the following: (a) Acute lesions were only observed when antigen was present and before antibody appeared in the circulation. (b) Healing of lesions was only observed (with one exception) when antigen had almost or completely disappeared from the circulation, usually with the appearance of antibody. (c) There was a correlation between the rapidity of evolution of the lesions and the rapidity with which the antigen disappeared from the circulation. (d) There was a rough correlation between the proportion of animals showing lesions and the proportion developing antibodies after the injection of a particular protein solution.

Binding of Antibodies onto the Thylakoid Membrane VII. Localization of Coupling Factor of Photophosphorylation in the Lamellar System of Chloroplasts from Antirrhinum majus

1982 ◽  
Vol 37 (3-4) ◽  
pp. 236-241 ◽  
Author(s):  
Alfons Radunz ◽  
Renate Meier
Keyword(s):  
Outer Surface ◽  
Thylakoid Membrane ◽  
Major Part ◽  
Coupling Factor ◽  
Antirrhinum Majus ◽  
Antigenic Determinants ◽  
Β Subunit ◽  
Lamellar System ◽  
Γ Subunit ◽  
Intact Chloroplasts

Abstract The maximal binding of antibodies against the subunits of the coupling factor o f photo­ phosphorylation onto the ultrasonic sediment and to stroma-freed chloroplasts of Antirrhinum majus was determined. Different surfaces o f the thylakoid membrane are accessible to antibodies in both chloroplast preparations. Stroma-freed chloroplasts bind antibodies only at the outer surface, which is directed towards the stroma in intact chloroplasts. In the ultrasonic sediment, which is the product of ultrasonication and centrifugation, as was demonstrated by electronmicroscopy, the major part of the surface directed towards the inside of the thylakoids is accessible.Both chloroplasts preparations are able to bind different amounts of antibodies to the five sub­ units of the coupling factor. While antigenic determinants of all five subunits are present on the surface directed to the outside, the δ-components seems to be lacking on the surface directed towards the inside. The α- and ε-component bind on both surfaces nearly the same amount of antibodies. Antigenic determinants of the β-subunit seem to exist in higher concentrations on the surface towards the outside and those of the γ-subunit on the surface towards the inside.From the serological investigations it follows that coupling factor molecules span the thylakoid membrane from the outside to the inside. They are not only present in those parts of the thylakoid membrane, exposed in intact chloroplasts to the stroma, but also in stacked thylakoids. The differences between the results of other authors and the present results are discussed.

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