Binding of Antibodies onto the Thylakoid Membrane II. Distribution of Lipids and Proteins at the Outer Surface of the Thylakoid Membrane

1977 ◽  
Vol 32 (7-8) ◽  
pp. 597-599 ◽  
Author(s):  
Alfons Radunz

Abstract The number of antibody molecules which stroma-freed chloroplasts can bind out of the mono-specific antisera to monogalactosyl diglyceride, tri-and digalactosyl diglyceride, sulfoquinovosyl diglyceride, phosphatidyl glycerol, sitosterol, plastoquinone, lutein and neoxanthin was determined. This number was compared to the number of antibody molecules which stroma-freed chloroplasts can maximally bind. The result is that the antibodies to the individual lipids cover at most 17 per cent of the accessible thylakoid membrane surface. From a serum which contains both antibodies to the proteins and lipids of the thylakoid membrane, not more antibody molecules are bound than from a serum to the proteins. This means that antibodies to proteins are able to cover up the entire accessible surface of the thylakoids whereas a mixture of antibodies to the lipids, listed above, cover only one forth of the surface. Consequently, antibodies which are bound to proteins can cover up the lipid areas entirely and in turn antibodies which are bound to lipids cover up parts of the protein areas. From this it follows that the portion of the surface, which is made up by lipids must be considerably smaller than 24 per cent. Furthermore, it follows from these experiments that the lipid areas are small and that lipids probably only fill up the gaps between the protein molecules.

1979 ◽  
Vol 34 (12) ◽  
pp. 1199-1204 ◽  
Author(s):  
Alfons Radunz

Binding of antibodies to proteins and lipids onto fragments of the thylakoid membrane was stu­died and compared with the binding of antibodies by stroma-freed chloroplasts. The membrane fragments were prepared from stroma-free chloroplasts by ultrasonication and fractional centrifugation. The fragments have an average diameter of 200 A. Their thickness corresponds to that of the thylakoid membrane. The membrane fragments adsorb out of an antiserum to lipids approximately the same amount of antibodies as out of an antiserum to proteins. In comparison to this, stroma-free chloroplasts bind 4 times more antibodies to proteins than to lipids. From this it follows that the major part of the lipids is located in the membrane surface which is directed towards the inside or is located inside the membrane. As the chemical analysis has shown these results are not caused by an altered chemical composition of the membrane fragments. Despite the fact that membrane proteins bind considerably less protein antibodies than stroma-free chloroplasts, the antibody binding in membrane fragment might be considerably increased for certain proteins such as a polypeptide with an apparent molecular weight 24000 and cytochrome f. Antibodies to the major components of the lipid mixture, such as to monogalactosyl diglyceride, trigalactosyl diglyceride, sulfoquinovosyl diglyceride and phosphatidyl glycerol are 3 to 4 times more bound by membrane fragments than by stroma-free chloroplasts. From these results it is concluded that the thylakoid membrane surface directed towards the inside is preponderently composed of lipids whereas the surface directed towards the outside consists only by 10 to 15% of lipids.


1980 ◽  
Vol 35 (11-12) ◽  
pp. 1024-1031 ◽  
Author(s):  
Alfons Radunz

The maximal binding of antibodies out of monospecific antisera to proteins and lipids onto three different chloroplast preparations is compared. In these preparations different parts of the thylakoid membrane surface are accessible to antibodies. Whereas stroma-freed chloroplasts bind antibodies only at the outer surface, also the inner membrane surface is exposed in the two chloroplast preparations which were obtained by ultrasonication and subsequent fractionating centrifugation. In the ultrasonic sediment the inner surface is prevailing. In the ultrasonic supernatant antibodies do not only react with the inner and outer surface but also with con­siderable parties of the interior of the thylakoid membrane. It was found that the thylakoid membrane surface exposed to the stroma consists preponder­antly of proteins whereas the surface directed towards the interior of the thylakoids consists mainly of lipids. All proteins involved in electron transport such as ferredoxin, ferredoxin- NADP+-reductase, plastocyanin, cytochrome f and the coupling factor of photophosphorylation are detectable in the outer surface. The molecules of the coupling factor span the thylakoid membrane from the outside to the inside. They hinder the binding of antibodies to mono- galactosyl diglyceride and to a polypeptide with the apparent molecular weight 24000. The polypeptide 24000 is a major component of the membrane proteins, and is detected on the outer and inner surface of the membrane. The major part of this polypeptide, however, is located in the interior of the thylakoid membrane. The lipid mixture has a different composition in the outer surface than on the inside face of the membrane. The sulfoquinovosyl diglyceride and the phosphatides, phosphatidyl glycerol, phosphatidyl cholin and phosphatidyl inositol are stronger concentrated in the outer surface than in the inside face. The neutral monogalactosyl diglyceride and di- or trigalactosyl diglyceride, however, occur in the inner surface in higher concentrations than in the outer surface. The major part of the sulfoquinovosyl diglyceride and of the monogalactosyl diglyceride are located in the interior of the membrane.


1975 ◽  
Vol 30 (7-8) ◽  
pp. 484-488 ◽  
Author(s):  
Alfons Radunz

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.


1978 ◽  
Vol 33 (11-12) ◽  
pp. 941-947 ◽  
Author(s):  
Alfons Radunz

Antisera to the phosphatides phosphatidyl inositol and phosphatidyl choline as well as to the xanthophylls violaxanthin and zeaxanthin were obtained by immunization of rabbits. The phosphatidyl choline antiserum did not give cross reactions with thylakoid membrane glycolipids, nor with phosphatidyl glycerol and phosphatidyl inositol. The antiserum to phosphatidyl inositol also did not react with the glycolipids and phosphatidyl choline. However, a cross reaction with phosphatidyl glycerol was observed. Exhaustion tests revealed, that the phosphatidyl inositol antiserum was fully neutralized with the homologous phosphatide, but not with phosphatidyl glycerol. Antibodies to the phosphatides phosphatidyl inositol and phosphatidyl choline react with stroma- freed, not swellable chloroplasts of Antirrhinum majus only in a monovalent manner as demon­strated by means of the anti-y-globuline consumption test. Sonicated and spun down chloroplasts, however, are agglutinated. Also an antiserum to phosphatidyl inositol treated with exhausting quantities of phosphatidyl glycerol agglutinates this ultrasonic sediment. This type of chloroplast preparation binds approximately double the amount of antibodies to phosphatides. Small membrane fragments of the ultrasonic supernatant in which the outer as well as the inner surface of the thylakoid membrane is accessible to antibodies, are not precipitated. They react with phosphatide antisera just like untreated chloroplasts. Chloroplasts of Antirrhinum majus and of Nicotiana tabacum, which were isolated in Tris buffer and whose thylakoids are swellable and exhibit high Hill reaction rates, were agglutinated by the phosphatide antisera. The antisera to violaxanthin and zeaxanthin gave with the described chloroplast preparations the same reactions as the phosphatide antisera. From this it is concluded that antigenic determinants of the phosphatides and xanthophylls are located in the surface of the thylakoid membrane directed towards the outside. Quantitative investigations on the maximal binding of antibodies showed that 1g stromafreed not swellable chloroplasts of Antirrhinum majus bind maximally 0.06 g antibodies to phosphatidyl choline and approximately four times the amount of antibodies to phosphatidyl inositol. Antibodies to the xanthophylls were bound in an amount of 0.04 to 0.09 g. The order of magnitude of these values fits the values of the maximal binding of antibodies to the glycolipids and also to proteins, which participate in photosynthetic electron transport. Onto the outer surface of the thylakoid membrane, accessible to antibodies, 1 g of antibodies can be maximally bound by 1 g chloroplasts.


Membranes ◽  
2022 ◽  
Vol 12 (1) ◽  
pp. 79
Author(s):  
Miroslav Kukučka ◽  
Nikoleta Kukučka Stojanović

Commercial nanofiltration membranes of different molecular weight cut-offs were tested on a pilot plant for the exploration of permeation nature of Ca, Mg, Mn, Fe, Na and ammonium ions. Correlation of transmembrane pressure and rejection quotient versus volumetric flux efficiency on nanofiltration membrane rejection and permeability behavior toward hydrated divalent and monovalent ions separation from the natural groundwater was observed. Membrane ion rejection affinity (MIRA) dimension was established as normalized TMP with regard to permeate solute moiety representing pressure value necessary for solute rejection change of 1%. Ion rejection coefficient (IRC) was introduced to evaluate the membrane rejection capability, and to indicate the prevailed nanofiltration partitioning mechanism near the membrane surface. Positive values of the IRC indicated satisfactory rejection efficiency of the membrane process and its negative values ensigned very low rejection affinity and high permeability of the membranes for the individual solutes. The TMP quotient and the efficiency of rejection for individual cations showed upward and downward trends along with flux utilization increase. Nanofiltration process was observed as an equilibrium. The higher the Gibbs free energy was, cation rejection was more exothermic and valuably enlarged. Low Gibbs free energy values circumferentially closer to endothermic zone indicated expressed ions permeation.


1977 ◽  
Vol 32 (3-4) ◽  
pp. 271-280 ◽  
Author(s):  
Georg H. Schmid ◽  
Alfons Radunz ◽  
Wilhelm Menke

Abstract A monospecific antiserum to cytochrome f agglutinates stroma-free swellable chloroplasts from tobacco and Antirrhinum. Consequently, antigenic determinants towards which the antiserum is directed are located in the outer surface of the thylakoid membrane. The antiserum inhibits linear photosynthetic electron transport. Just as described earlier for the antiserum to polypeptide 11000 this inhibition develops in the course of the light reaction. Ultrasonication in the presence of anti­ serum abolishes the light requirement and the maximal inhibition of the electron transport reaction is immediately observed. Electron transport in chloroplasts from a tobacco mutant which ex­ hibits only photosystem I-reactions is also inhibited by the antiserum. No time lag in the light for the onset of inhibition is observed with these chloroplasts. As chloroplasts of this mutant have only single unfolded thylakoids it appears that light might preponderantly open up partitions. If the light effect is interpreted in this way, cytochrome f should be located in the partition regions but nevertheless in the outer surface of the thylakoid membrane. However, a rearrangement of molecules in the membrane in the light by which the accessibility of cytochrome f is changed can­ not be excluded. The inhibition of linear electron transport by the antiserum is approximately 50 per cent and can only be increased to 75% upon the addition of antibodies to plastocyanin. The inhibition by the antiserum to cytochrome f as well as the combined inhibition by the antisera to cytochrome f and plastocyanin can be by-passed by DCPiP. It appears that cytochrome f and plastocyanin cannot be connected in series in the electron transport chain but are both closely associated in the thylakoid membrane. PMS-mediated cyclic photophosphorylation in chloroplasts from wild type tobacco and the tobacco mutant NC95 is only inhibited if the chloroplasts are sonicated in the presence of anti­ serum. If one disregards, that ultrasonication might cause reaction artifacts, it is thinkable that the cytochrome f, involved in the PMS-mediated cyclic photophosphorylation reaction, might be located inside the membrane.


Symmetry ◽  
2020 ◽  
Vol 12 (8) ◽  
pp. 1271
Author(s):  
Jiri Neustupa ◽  
Yvonne Nemcova

Calcifying marine green algae of genus Halimeda have siphonous thalli composed of repeated segments. Their outer surface is formed by laterally appressed peripheral utricles which often form a honeycomb structure, typically with varying degrees of asymmetry in the individual polygons. This study is focused on a morphometric analysis of the size and symmetry of these polygons in Mediterranean H. tuna. Asymmetry of surface utricles is studied using a continuous symmetry measure quantifying the deviation of polygons from perfect symmetry. In addition, the segment shapes are also captured by geometric morphometrics and compared to the utricle parameters. The area of surface utricles is proved to be strongly related to their position on segments, where utricles near the segment bases are considerably smaller than those located near the apical and lateral margins. Interestingly, this gradient is most pronounced in relatively large reniform segments. The polygons are most symmetric in the central parts of segments, with asymmetry uniformly increasing towards the segment margins. Mean utricle asymmetry is found to be unrelated to segment shapes. Systematic differences in utricle size across different positions might be related to morphogenetic patterns of segment development, and may also indicate possible small-scale variations in CaCO3 content within segments.


1977 ◽  
Vol 32 (9-10) ◽  
pp. 817-827 ◽  
Author(s):  
Friederike Koenig ◽  
Wilhelm Menke ◽  
Alfons Radunz ◽  
Georg H. Schmid

Abstract Three polypeptide fractions with the apparent molecular weight 66 000 were isolated from stroma-freed Antirrhinum chloroplasts which were solubilized with dodecyl sulfate. Antisera to these fractions affect electron transport in distinctly different ways. For the characterization of the three antisera photochemical reactions of chloroplast preparations with artificial electron donors and acceptors as well the analysis of fluorescence rise curves were used. Antiserum 66 000 PSI-96 inhibits electron transport apparently on the acceptor side of photosystem I, provided the antibodies are adsorbed onto the outer surface of the thylakoid membrane. Antiserum 66 000 PSI-88 probably acts directly on the reaction centre I or on its immediate vicinity, if the antibodies are adsorbed at the inner surface of the thylakoid membrane. Antiserum 66 000 PSII-42 inhibits electron trans­ port in the region of photosystem II. The antigen towards which the antiserum is directed appears to belong to the reaction centre II, as also in the condition of high inhibition degrees, the fluorescence intensity remains unchanged. The antigenic determinants are located at the outer surface of the thylakoid membrane.


Sign in / Sign up

Export Citation Format

Share Document