Stoichiometry of proton uptake in isolated pea chloroplasts under different light intensities

The number of protons released inside the chloroplast thylakoids per electron which is transferred through the electron transport chain (H⁺/e^– ratio) was measured in isolated pea chloroplasts at pH 6.0 under continuous illumination and with methyl viologen as an electron acceptor. At saturating light intensity (200 W · m^–2) (strong light) the H⁺/e^– ratio was 3. At low intensity (0.9 W · m^–2) (weak light) the H⁺/e^– ratio was 2 with dark-adapted chloroplasts, but it was close to 3 with chloroplasts that were preilluminated with strong light. It is shown that the presence of azide in the reaction mixture leads to errors in the determination of the H⁺/e^– ratio due to underestimation of the initial rate of H⁺ efflux on switching off the light. To explain the above data, we assume that transformation of the electron transport chain occurs during illumination with strong light, namely, the Q cycle becomes operative.     

Binding of antibodies in liposomes to extracellular matrix antigens

We have incorporated antibodies against fibronectin or laminin into liposomes and studied their interaction with insoluble forms of these antigens. The antibodies, after modification by palmitoylchloride, were incorporated into the lipid bilayer by the cholate dialysis method. The antibodies in the liposomes recognized their specific antigen with little reaction to the alternative attachment protein or to albumin (less than 2%). The binding of antibody-containing liposomes to insoluble antigen was inhibited by soluble antibodies to the respective antigens but not by antibodies to other antigens. The affinity constant of the liposome-antibody complex with the antigen was estimated at 1-10 X 10(-9) M liposomes. Thus, antibodies in liposomes retain their reactivity and specificity, and the reaction constant is comparable to that observed for immune complexes.     

The absence of non-local conformational changes in OR3 operator DNA on complexing with the cro repressor

The Interaction of the cro protein of lambda phage with a synthetic OR3 operator having 17 base pairs in length and with its 9 bp fragment has been studied using the circular dichroism (CD) method. In both cases, a considerable change in the CD of the samples was found in the region 260-300 nm upon the addition of the cro protein. The stoichiometry obtained by the CD titration was identical for OR3 and its 9 bp fragment: one duplex per dimeric cro. NaCl addition makes the complexes dissociate so that the 9 bp fragment becomes free at [NaCl] greater than 0.2 M while the whole OR3 becomes free at [NaCl] greater than 0.5 M. The CD spectra of both the free duplexes show a typical B-form conservative pattern with a positive CD band (270 nm) and a negative one (250 nm). The specific complexing of both the duplexes results in a substantial CD depression in the positive band. The most pronounced effect occurs at 280 nm. This spectral change is quite distinct from those in the B to A transition and in the non-cooperative winding of the DNA within the B-family of forms. The interaction of the cro protein with the non-operator DNAs, calf thymus DNA and a synthetic 10 bp duplex, reveals no visible CD changes at all. An inference is drawn that the CD change in the specific complexes is mainly due to the induced CD in tyr-26 upon its interaction with a specific base pair in the operator or its fragment, the operator DNA conformation being conserved in a B-like form as a whole.(ABSTRACT TRUNCATED AT 250 WORDS)     

Peculiarities of the B to A transition of the lambda phage regulatory site OR3 and of its fragment

Conformations of the synthetic deoxyoligonucleotide 17 base pairs long, which is an OR3 operator of lambda phage, and of its 9-b.p. fragment were studied by the circular dichroism method (CD). The regions of stability of the double-stranded state were determined for these duplexes. A comparison of the CD spectra for these oligonucleotides with the CD for a lengthy DNA showed the conformation of these short DNA pieces to belong to the B-family. A cooperative change in the CD spectra is observed in trifluoroethanol (TFE) solutions at a TFE concentration specific for each oligonucleotide, which is supposed to stem from a B to A transition. The length of the fragment was found to affect the ability for the B-A transition. The transition into the A form is hindered by 13% TFE for the short 9-nucleotide in comparison with the 17-nucleotide. We suggest that this is due to the B form stabilization by terminal base pairs (B-phility of the ends).     

Aminoacyl-tRNA synthetases: inter-site interaction as a possible proofreading mechanism

Smooth muscle cell energetics of taenia caeci during relaxation, activity and maximal contraction were investigated using ³¹P-NMR. In relaxed muscle obtained in calcium-free medium, [ATP], [phosphocreatine] and [sugar phosphate] were 4.4 mM, 7.7 mM and 2.8 mM, respectively. There was only a small difference in the energetics of spontaneously active and maximally contracted muscles, but under both conditions substantial changes occurred as compared with relaxed muscles. The internal pH in relaxed muscle was found to be 7.05, which acidified to 6.5 during contraction. The level of sugar phosphates was found to be not a limiting factor in energetics.     

Some theoretical aspects of protein coevolution in the ecosystem “phage-bacteria” II. The deterministic model of microevolution

The deterministic model of microevolutionary dynamics of “phagebacteria” ecosystem is analyzed. Primary (and after all decisive) events that determine the dynamics are direct interactions between bacterial reception and viral adsorption proteins. Structure of the model is that under real parameters of adsorption, lysis and reproduction each separate (ith) stage of microevolution comes to end with total lysis of ith population of bacteria by ith population of phage. It is shown however that in the course of joint microevolution both populations pass over some critical sizes when a new pair of antagonistic strains arises with certainty from mutations. As a result it is easy to visualize and simulate by computer the process of successive fixations of such pairs of mutants. This coevolution is the original example of a locally adaptive but globally undirected process which is characterized also by: (1) constant average rate, (2) neutrality of mutations at the moment of their emergence and during the period of “anticipation” of ecological changes, (3) pure adaptivity of the same mutations at the moment of proper fixation and (4) “intrinsic origin” (from the ecosystem dynamics itself) of selective constraints.     

Unusual genome organization of the marine invertebrate Rapana thomasiana Grosse (Gastropoda)

The genome organization of the marine snail Rapana thomasiana Grosse (Gastropoda), genome size 2.7 pg, was studied by reassociation kinetics, S₁-nuclease assay, and restriction enzyme analysis. The slow-reassociating (single-copy) fraction represented only 21% of the genome. The average length of 80% of the single-copy sequences was less than 700 bp and the remaining 20% no longer than 1,400 bp. Longer stretches of unique DNA were not observed. The genome contained an unusually high percent-age of inverted repeats: at standard fragment length the zero-time binding fraction amounted to 25% of the genome. Foldback structures ranging from 200 bp to more than 10 kb were observed after S₁-nuclease treatment. They were randomly distributed throughout at least 85% of the genome, and the spacings between them were estimated to be about 1,600 bp on the average. The middle-repetitive DNA (45% of the genome) contained two kinetic components, repeated 430 and 65,000 times per genome, respectively. It was found that the majority of the repetitive sequences are about 300 bp long. Longer repeats (about 2,000 bp) were also observed, comprising a small portion of the genome. The inverted repeats, the middle-repetitive, and the singly-copy sequences were fully interspersed in the genome, thus indicating that R. thomasiana DNA is not organized in either the Xenopus or the Drosophila pattern type. — R. thomasiana is the only mollusc so far in which a satellite DNA has been found. It is organized in tandem repeats of 1,460 bp with a very complex organization but a low degree of divergence.