Aliphatic 1H and 13C resonance assignments for the 26-10 antibody VL domain derived from heteronuclear multidimensional NMR spectroscopy

Summary Extensive ¹H and ¹³C assignments have been obtained for the aliphatic resonances of a uniformly ¹³C-and ¹⁵N-labeled recombinant V_L domain from the anti-digoxin antibody 26-10. Four-dimensional triple resonance NMR data acquired with the HNCAHA and HN(CO)CAHA pulse sequences [Kay et al. (1992) J. Magn. Reson., 98, 443–450] afforded assignments for the backbone H^N, N, H and C resonances. These data confirm and extend H^N, N and H assignments derived previously from three-dimensional ¹H-¹⁵N NMR studies of uniformly ¹⁵N-labeled V_L domain [Constantine et al. (1992), Biochemistry, 31, 5033–5043]. The identified H and C resonances provided a starting point for assigning the side-chain aliphatic ¹H and ¹³C resonances using three-dimensional HCCH-COSY and HCCH-TOCSY experiments [Clore et al. (1990), Biochemistry, 29, 8172–8184]. The C and C chemical shifts are correlated with the V_L domain secondary structure. The extensive set of side-chain assignments obtained will allow a detailed comparison to be made between the solution structure of the isolated V_L domain and the X-ray structure of the V_L domain within the 26–10 Fab.     

Metal Cations Defibrillize the Amyloid Beta-Protein Fibrils

Amyloid beta-protein (A) is the major constituent of amyloid fibrils composing -amyloid plaques and cerebrovascular amyloid in Alzheimer's disease (AD). We studied the effect of metal cations on preformed fibrils of synthetic A by Thioflavin T (ThT) fluorescence spectroscopy and electronmicroscopy (EM) in negative staining. The amount of cross beta-pleated sheet structure of A 1–40 fibrils was found to decrease by metal cations in a concentration-dependent manner as measured by ThT fluorescence spectroscopy. The order of defibrillization of A 1–40 fibrils by metal cations was: Ca²⁺ and Zn²⁺ (IC₅₀ = 100 M) > Mg²⁺ (IC₅₀ = 300 M) > Al³⁺ (IC₅₀ =1.1 mM). EM analysis in negative staining showed that A 1–40 fibrils in the absence of cations were organized in a fine network with a little or no amorphous material. The addition of Ca²⁺, Mg²⁺, and Zn²⁺ to preformed A 1–40 fibrils defibrillized the fibrils or converted them into short rods or to amorphous material. Al³⁺ was less effective, and reduced the fibril network by about 80 % of that in the absence of any metal cation. Studies with A 1–42 showed that this peptide forms more dense network of fibrils as compared to A 1–40. Both ThT fluorescence spectroscopy and EM showed that similar to A 1–40, A 1–42 fibrils are also defibrillized in the presence of millimolar concentrations of Ca²⁺. These studies suggest that metal cations can defibrillize the fibrils of synthetic A.     

The Use of the [13C]/[12C] Ratio for the Assay of the Microbial Oxidation of Hydrocarbons

The study deals with a comparative analysis of the relative abundances of the carbon isotopes ¹²C and ¹³C in the metabolites and biomass of the Burkholderia sp. BS3702 and Pseudomonas putida BS202-p strains capable of utilizing aliphatic (n-hexadecane) and aromatic (naphthalene) hydrocarbons as sources of carbon and energy. The isotope compositions of the carbon dioxide, biomass, and exometabolites produced during the growth of Burkholderia sp. BS3702 on n-hexadecane (¹³C = –44.6 ± 0.2) were characterized by the values of ¹³C_{CO 2} = –50.2 ± 0.4, ¹³C_biom = –46.6 ± 0.4, and ¹³C_exo = –41.5 ± 0.4, respectively. The isotope compositions of the carbon dioxide, biomass, and exometabolites produced during the growth of the same bacterial strain on naphthalene (¹³C = –21 ± 0.4) were characterized by the isotope effects ¹³C_{CO 2} = –24.1 ± 0.4, ¹³C_biom = –19.2 ± 0.4, and ¹³C_exo = –19.1 ± 0.4, respectively. The possibility of using the isotope composition of metabolic carbon dioxide for the rapid monitoring of the microbial degradation of petroleum hydrocarbons in the environment is discussed.     

Expression pattern of voltage-dependent calcium channel α1 and β subunits in adrenal gland of N-type Ca2+ channel α1B subunit gene-deficient mice

The Ca²⁺ channel _1B subunit is a pore-forming component capable of generating N-type Ca²⁺ channel activity. Although N-type Ca²⁺ channel plays a role in a variety of neuronal functions, _1B-deficient mice exhibit normal life span without apparent abnormalities of behavior, histology or plasma norepinephrine level, presumably owing to compensation by some other Ca²⁺ channel ₁ or subunit. In this study, we studied the levels of _1A, _1C, _1D, _1E, ₁, ₂, ₃ and ₄ mRNAs in adrenal gland of _1B-deficient mice. The _1A mRNA in homozygous mice was expressed at higher level than in wild or heterozygous mice, but no difference in the expression levels of _1C, _1D, _1E, ₁, ₂, ₃ and ₄ was found among wild, heterozygous and homozygous mice. The protein level of _1A in homozygous mice was also expressed at higher level than in wild or heterozygous mice. To examine whether increased expression is induced by cis-regulatory element within 5-upstream region of _1A gene, we examined lacZ expression in _1B-deficient × _1A6.3-lacZ mice (carrying a 6.3-kb 5-upstream fragment of _1A gene fused to E. coli lacZ reporter gene), which express lacZ in medullar chromaffin cells, but not in cortex. The levels of lacZ expression in homozygous _1B-deficient × _1A6.3-lacZ mice were higher than in wild or heterozygous mice. Therefore, a possible explanation of the normal behavior and plasma norepinephrine level of _1B-deficient mice is that compensation by _1A subunit occurs and that 6.3-kb 5-upstream region of _1A gene contains enhancer cis-element(s) for compensation in adrenal medulla chromaffin cells. (Mol Cell Biochem 271: 91–99, 2005)     

A tracked approach for automated NMR assignments in proteins (TATAPRO)

A novel automated approach for the sequence specific NMR assignments of ¹H^N, ¹³C, ¹³C, ¹³C/¹H and ¹⁵N spins in proteins, using triple resonance experimental data, is presented. The algorithm, TATAPRO (Tracked AuTomated Assignments in Proteins) utilizes the protein primary sequence and peak lists from a set of triple resonance spectra which correlate ¹H^N and ¹⁵N chemical shifts with those of ¹³C, ¹³C and ¹³C/¹H. The information derived from such correlations is used to create a `master_list' consisting of all possible sets of ¹H^N _i, ¹⁵N_i, ¹³C _i, ¹³C _i, ¹³C_i/¹H _i, ¹³C _i–1, ¹³C _i–1 and ¹³C_i–1/ ¹H _i–1 chemical shifts. On the basis of an extensive statistical analysis of ¹³C and ¹³C chemical shift data of proteins derived from the BioMagResBank (BMRB), it is shown that the 20 amino acid residues can be grouped into eight distinct categories, each of which is assigned a unique two-digit code. Such a code is used to tag individual sets of chemical shifts in the master_list and also to translate the protein primary sequence into an array called pps_array. The program then uses the master_list to search for neighbouring partners of a given amino acid residue along the polypeptide chain and sequentially assigns a maximum possible stretch of residues on either side. While doing so, each assigned residue is tracked in an array called assig_array, with the two-digit code assigned earlier. The assig_array is then mapped onto the pps_array for sequence specific resonance assignment. The program has been tested using experimental data on a calcium binding protein from Entamoeba histolytica (Eh-CaBP, 15 kDa) having substantial internal sequence homology and using published data on four other proteins in the molecular weight range of 18–42 kDa. In all the cases, nearly complete sequence specific resonance assignments (> 95%) are obtained. Furthermore, the reliability of the program has been tested by deleting sets of chemical shifts randomly from the master_list created for the test proteins.     

An H-2 haplotype possibly derived by crossing-over between the (A α A β ) duplex and the E βlocus

The B10.STA62 strain carries the H-2 ^w27 haplotype derived from a wild mouse captured in the vicinity of Ann Arbor, Michigan. Products of two class II loci composing this haplotype, A and A , are serologically, biochemically (by tryptic peptide mapping), and functionally indistinguishable from products controlled by the A ^b and A ^/b genes of the B10.A(5R) strain. In contrast, the polypeptide chain controlled by the third class II locus, E , is different from that controlled by the E ^/b gene. This E ^/w27 chain lacks an antigenic determinant present on the E^b molecule and carries determinants lacking on the E^b molecule, the E ^/b and E ^/w27 peptide maps differ in at least six peptides, and cytotoxic T cells specific for the E ^b chains do not react with B10.STA62 target cells. This great difference between the E ^/b and E ^/w27 chains suggests that the corresponding genes have not been derived from one another by a direct mutational conversion; instead, H-2 ^w27 appears to be a recombinant haplotype derived by crossing-over between the A A duplex and the E locus. This is the first recombinant discovered separating these class II loci.     

Growth of the Thermophilic Bacterium Geobacillus uralicus as a Function of Temperature and pH: An SCM-Based Kinetic Analysis

The synthetic chemostat model (SCM), originally developed to describe nonstationary growth under widely varying concentrations of the limiting substrate, was modified to account for the effects of nontrophic factors such as temperature and pH. The bacterium Geobacillus uralicus, isolated from an ultradeep well (4680 m), was grown at temperatures ranging from 40 to 75°C and at pH varying from 5 to 9. The biomass kinetics was reasonably well described by the SCM, including the phase of growth deceleration observed in the first hours after a change in the cultivation temperature. At an early stage of batch growth in a neutral or alkalescent medium, bacterial cells showed reversible attachment to the glass surface of the fermentation vessel. The temperature dependence of the maximum specific growth rate (_m) was fitted using the equation _m = Aexp(T)/{1 + expB[1 – C/(T + 273)]}, where A, , B, and C are constants. The maximum specific growth rate of 2.7 h^–1 (generation time, 15.4 min) was attained on a complex nutrient medium (peptone and yeast extract) at 66.5°C and pH 7.5. On a synthetic mineral medium with glucose, the specific growth rate declined to 1.2 h^–1, and the optimal temperature for growth decreased to 62.3°C.     

Linked genetic markers of the rabbit κ light chain are not linked to the Tcr β chain genes

In order to investigate linkage, we used serum allotypes of the two rabbit C isotypes and restriction fragment length polymorphisms (RFLPs) of the genes for V , C , and T-cell receptor C . The inheritance of these genetic markers was studied through backcross and F₂ matings. Southern analysis and hybridization of genomic DNA with a C probe detected a 5 kb Pst I fragment linked to expression of the K2bas1 allotype and the presence of the 1b ^bas gene and a 6.6 kb Pst I fragment linked to the expression of the K1b9 allotype, the presence of the 2 ^bas2 gene and lack of expression of the K2bas1 allotype. A V probe detected a 1.3 kb Eco RI fragment linked to the presence of the 1b ^bas gene and expression of the K2bas1 allotype. In contrast, the 9 or 14 kb Eco RI RFLP (C ^a or C ^b) detected with a Tcr chain probe segregated independently from C allotypes and RFLPs. It has previously been found that C and C are also unlinked in man, whereas in the mouse they are linked at a distance of 8 centimorgans.     

Mutants in yeast affecting ethidium bromide induced rho- formation and their effects on transmission and recombination of mitochondrial genes.

Summary A series of mutants called ebi, less inducible by ethidium bromide than the parental strain for the ^+– mutation have been isolated after E.M.S. mutagenesis. Some of the ebi mutants also show an important accumulation of ^– cells, in the absence of ethidium bromide. Ebi mutations are nuclearly inherited as shown by meiotic segregation. The effects of these mutants on the transmission and recombination of mitochondrial genes among the diploid progeny of crosses have been studied. Some of the ebi mutants show a non coordinated transmission of the oli1 mitochondrial marker with respect to other mitochondrial markers unexpected for homosexual crosses. This bias which is independent from will be discussed in relation to the segregation and recombination. No significant decrease of the frequency of recombinants has been detected.Abbreviations E.B. Ethidium bromide - E.M.S. Ethyl méthane sulfonate - C^S/C^R Allelic forms of the rib 1 locus conferring chloramphenicol sensitivity/resistance - E^S/E^R Allelic forms of the rib 3 locus conferring erythromycine sensitivity/resistance - O^R/O^R Allelic forms of the oli 1 locus conferring oligomycin sensitivity/resistance - P^S/P^R Allelic forms of the par 1 locus conferring paromomycine sensitivity/resistance - D^S/D^R Allelic forms of the diu 1 or diu 2 loci conferring diuron sensitivity/resistance - C^S/C^R Allelic forms of the mitochondrial locus - ⁺ grande or respiratory competent cells - ^– petite or cytoplasmic respiratory deficient cells     

Effect of microbial growth on pore entrance size distribution in sandstone cores

Summary The in situ growth of microorganisms in Berea sandstone cores preferentially plugged the larger pore entrances. The largest pore entrance sizes after microbial plugging ranged from 20 to 38 m, compared with 59 to 69 m before plugging. The pore entrance size distribution of plugged cores was shifted to smaller sizes. A mathematical model based on Poiseuille's equation was found to adequately predict permeability reductions (greater than 90%) caused by microbial growth in the large pore entries.Nomenclature Q volumetric flow rate (L ₃/t) - C orifice constant (dimensionless) - A cross-sectional area (L ₂) - g gravity (L/t ²) - h pieziometric head (L) - _s transmittivity (L ²) - R _e Reynolds number (dimensionless) - a constant (dimensionless) - density (M/L ₃) - viscosity (M/Lt) - d diameter (L) - L length (L) - P pressure change (M/L ₂)     

Biological reasons for the unsuitability of growth parameters and indices for comparing fish growth

Parameters k and L in the Von Bertalanffy equation and their derivatives =kL, = logk + 2logL; as well as slopes b_L/t of the regression L=at^b (t, years), relative increments (CI=[L₂–L₁]/L₁), specific growth rates (C_v=InL₂–InL₁), growth characteristics (Clh = CvxL₁) and growth constants (Clt=Cvx[t₂ + t₁]/2) were analyzed. A total of 121 bream Abramis brama stocks in the first 10 years of life were studied. At the same real growth rate (the average absolute linear annual increments, mm year^–1) the values of k, L, , , b_L/t, CI, Cv and Clt in different stocks vary within almost the whole range. The main reason is the natural process of growth self-regulation: the relation between the average body lengths in the first year (L₁) and the relative growth rates (slopes b_L/t) is negative (b_L/t = exp[0.1183–0.0053L₁], r=0.76). The above relation defines 4 principal types of the Ford-Walford lines. Thirty four percent of the stocks have rather steep slopes of the lines and even parallel the absolute slope of 45°, so the L values of these stocks have no biological significance. The authors recommend a simple and more precise, from a biological point of view, approach for comparing fish population growth rates.     

13C discrimination during CO2 assimilation by the terrestrial biosphere

Estimates of the extent of the discrimination against¹³CO₂ during photosynthesis (_A) on a global basis were made using gridded data sets of temperature, precipitation, elevation, humidity and vegetation type. Stomatal responses to leaf-to-air vapour mole fraction difference (D, leaf-to-air vapour pressure difference divided by atmospheric pressure) were first determined by a literature review and by assuming that stomatal behaviour results in the optimisation of plant water use in relation to carbon gain. Using monthly time steps, modelled stomatal responses toD were used to calculate the ratio of stomatal cavity to ambient CO₂ mole fractions and then, in association with leaf internal conductances, to calculate _A. Weighted according to gross primary productivity (GPP, annual net CO₂ asimilation per unit ground area), estimated _A for C₃ biomes ranged from 12.9 for xerophytic woods and shrub to 19.6 for cool/cold deciduous forest, with an average value from C₃ plants of 17.8. This is slightly less than the commonly used values of 18–20. For C₄ plants the average modelled discrimination was 3.6, again slightly less than would be calculated from C₄ plant dry matter carbon isotopic composition (yielding around 5). From our model we estimate that, on a global basis, 21% of GPP is by C₄ plants and for the terrestrial biosphere as a whole we calculate an average isotope discrimination during photosynthesis of 14.8. There are large variations in _A across the globe, the largest of which are associated with the precence or absence of C₄ plants. Due to longitudinal variations in _A, there are problems in using latitudinally averaged terrestrial carbon isotope discriminations to calculate the ratio of net oceanic to net terrestrial carbon fluxes.     

The influence of ethanol on the functional status of GABA(A) receptors

Gamma aminobutyric acid (GABA) is one of the main inhibitory neurotransmitters in the mammalian brain. Its effects are realized via GABA_A, GABA_B, and GABA_C receptors. GABA_A is the most abundant type of GABA receptors. It consists of six classes of subunits, , , , , , and . Acute and chronic exposures to ethanol are accompanied by changes in structure and function of GABA_A receptors. These changes may be a basis for altered behavior seen in alcoholism.     

Untersuchungen zum Polymorphismus der sauren Erythrocytenphosphatasen (E C 3.1.3.2.)

Zusammenfassung Die Bestimmungstechnik der sauren Erythrocytenphosphatasen wird eingehend beschrieben. Untersuchungen zur Formalgenetik bei 80 Familien mit 118 Kindern sowie zur Populationsgenetik und Phylogenetik werden mitgeteilt. Die Ergebnisse widersprechen nicht dem formalen Modell 3 Allele Ph^A, Ph^B, Ph^C an einem autosomalen Locus.

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The method for determination of red cell acid phosphatases is described in detail.Investigations on formal genetics (80 families with 118 children), population genetics and phylogenetics are communicated. The results agree with the assumption: 3 alleles Ph^A, Ph^B, and Ph^C at one autosomal locus.

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Wesentliche Teile dieser Arbeit werden von Fräulein U. Callsen als Dissertation der Medizinischen Fakultät der Universität Freiburg i. Br. vorgelegt.     

Production of hemoglobin C in the Moufflon (Ovis musimon pallas, 1811) and the Barbary sheep (Ammotragus lervia pallas, 1777) during experimental anemia: Amino acid composition of tryptic peptides from the βB and βC chains

A hemoglobin ^C chain has been detected in anemic Moufflon and in anemic Barbary sheep. Results of limited structural studies suggest that these ^C chains are closely related to the ^C chains which are produced in the domestic sheep and goat during experimental anemia and rather different from the ^C chain of certain nonanemic Barbary sheep. The structural variability among the chains of some nonanemic Caprini species is greater than that observed for the ^C chains of the same animals.This research was supported in part by United States Public Health Service Research grant HE-05168.     

Possible linkage between uncoiler chromosome Un 1 and mylase polymorphism Amy 2 loci

Summary A family has been investigated in which 7 from 12 members bear the uncoiler chromosome number 1 pair. In all these members with uncoiler chromosome 1 heterozygous amylase polymorphism variant Amy 2^A2^B has been estimated (with exception of one 3-month-old boy in whom the phenotype of amylase variant is not yet fully developed). This pedigree represents a typical double back-cross family which enables to suppose very probably the close linkage between both gene loci Un 1 and Amy 2 and also to assign Amy 2 locus to chromosome 1.

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Zusammenfassung Wir berichten über eine Familie, in welcher 7 unter insgesamt 12 untersuchten Familienmitgliedern ein asymmetrisches Chromosom des ersten Chromosompaares mit den sehr verlängerten langen Armen (uncoiler chromosome) gefunden wurde. Bei allen diesen Familienmitgliedern mit, uncoiler chromosome 1 wurde auch gleichzeitig die heterozygote Variante des Amylase-Polymorphismus Amy 2^A2^B gefunden (mit Ausnahme eines 3 Monate alten Knaben, bei welchem das Phenotyp der Amylase-Variante noch nicht völlig entwickelt wurde). Dieser Stammbaum stellt eine typische double back-cross-Familie vor, aus welcher mit großer Wahrscheinlichkeit die linkage zwischen beiden Genorten Un 1 und Amy 2 und damit auch die Lokalisation des Genortes Amy 2 auf dem Chromosom 1 hervorgeht.

     

Analysis of gas exchange in seedlings of Acer saccharum: integration of field and laboratory studies

In the field, photosynthesis of Acer saccharum seedlings was rarely light saturated, even though light saturation occurs at about 100 mol quanta m^-2 s^-1 photosynthetic photon flux density (PPFD). PPFD during more than 75% of the daylight period was 50 mol m^-2 s^-1 or less. At these low PPFD's there is a marked interaction of PPFD with the initial slope (CE) of the CO₂ response. At PPFD-saturation CE was 0.018 mol m^-2 s^-1/(l/l). The apparent quantum efficiency (incident PPFD) at saturating CO₂ was 0.05–0.08 mol/mol. and PPFD-saturated CO₂ exchange was 6–8 mol m^-2 s^-1. The ratio of internal CO₂ concentration to external (C _i /C _a ) was 0.7 to 0.8 except during sunflecks when it decreased to 0.5. The decrease in C _i /C _a during sunflecks was the result of the slow response of stomates to increased PPFD compared to the response of net photosynthesis. An empirical model, which included the above parameters was used to simulate the measured CO₂ exchange rate for portions of two days. Parameter values for the model were determined in experiments separate from the daily time courses being sumulated. Analysis of the field data, partly through the use of simulations, indicate that the elimination of sunflecks would reduce net carbon gain by 5–10%.List of symbols A measured photosynthetic rate under any set of conditions (mol m^-2 s^-1) - A _m (atm) measured photosynthetic rate at saturating PPFD, 350 l/l CO₂ and 21% (v/v) O₂ (mol m^-2 s^-1) - C constant in equation of Smith (1937, 1938) - C _a CO₂ concentration in the air (l/l) - C _i CO₂ concentration in the intercellular air space (l/l) - C _i ^/* C _i corrected for CO₂ compensation point, i.e., C _i -I ^*, (l/l) - CE initial slope of the CO₂ response of photosynthesis (mol m^-2 s^-1/(l/l)) - CEM CE at PPFD saturation - E transpiration rate (mmol m^-2 s^-1) - F predicted photosynthetic rate (mol m^-2 s^-1) - G leaf conductance to H₂O (mol m^-2 s^-1) - I photosynthetic photon flux density (mol m^-2 s^-1) - N number of data points - P _m predicted photosynthetic rate at saturating CO₂ and given PPFD (mol m^-2 s^-1) - P _ml predicted photosynthetic rate at saturating CO₂ and PPFD (mol m^-2 s^-1) - R _d residual respiratory rate (mol m^-2 s^-1) - T _a air temperature (°C) - T _l leaf temperature (°C) - V reaction velocity in equation of Smith (1937, 1938) - V _max saturated reaction velocity in equation of Smith (1937, 1938) - VPA vapor pressure of water in the air (mbar/bar) - VPD vapor pressure difference between leaf and air (mbar/bar) - X substrate concentration in equation of Smith (1937, 1938) - initial slope of the PPFD response of photosynthesis at saturating CO₂ (mol CO₂/mol quanta) - (atm) initial slope of the PPFD response of photosynthesis at 340 l/l CO₂ and 21% (v/v) O₂ (mol CO₂/mol quanta) - I ^* CO₂ compensation point after correction for residual respiration (l/l) - PPFD compensation point (mol m^-2 s^-1)     

Influence of Triterpene Glycosides of the Holothurian Cucumaria japonica on Root Growth in Cucumis sativus L. Seedlings

We have shown that triterpene glycosides of the Far Eastern edible holothurian Cucumaria japonica (monosulfated subfractions A ₂ and A ₄, disulfated subfraction A ₆, trisulfated subfraction A ₇, and the total fraction of monosulfated glycosides including subfractions A ₀, A ₁, A ₂, and A ₄) inhibited growth of the main root of Cucumis sativus seedlings. Cucumariosides were arranged as follows according to their inhibitory activity: subfraction A ₂ (ED₅₀ = 53.1 ± 3.6 g/ml) > total fraction of monosulfated glycosides (ED₅₀ = 127.4 ± 5.8 g/ml) > subfraction A ₄ (ED₅₀ = 346.5 ± 15.2 g/ml) > subfraction A ₆ (ED₅₀ = 375.7 ± 17.2 g/ml) > subfraction A ₇ (ED₅₀ = 539.4 ± 11.5 g/ml). The average length of the main root decreased with the increase of glycoside concentrations as a result of a growing amount of seedlings with low germination capacity and the absence of seedlings with high germination capacity.     

Polymorphism and Mendelian inheritance of photosystem II 23-kilodalton polypeptide

Soluble proteins from leaves of Nicotiana glauca Grah., N. langsdorffii Weinm., their reciprocal hybrids and amphiploid hybrid (N. glaucaxN. langsdorffii) were resolved by two-dimensional gel electrophoresis. Among a group of well-resolved polypeptides, in the isoelectric-point range of 5–5.5 and relative-molecular-mass (M_r) range of 18–23 kilodaltons (kDa), species-specific variation was observed. Polypeptides designated L and l are specific to N. langsdorffii, and G and g to N. glauca, while C is common to both species. Polypeptides L, G and C are localized in the chloroplasts and associated with thylakoid membranes. Polypeptide L is more acidic than polypeptide G, and both polypeptides have an M_r of 23 kDa. They were isolated from two-dimensional gels and their first 13 N-terminal amino-acid sequences were determined. These were found to be identical to the 13N-terminal amino acids of the photosystem II (PSII) 23-kDa polypeptide from spinach (T. Jansen et al. (1987) FEBS Lett. 216, 234–240) and, except for one change, to those from pea (R. Wales et al. (1989) Plant Molec. Biol., in press). Polypeptides G and L cross-react with antiserum against the PSII 23-kDa polypeptide from pea. Therefore, polypeptides G and L are extrinsic PSII 23-kDa polypeptides. They appear jointly and in equal amounts in the reciprocal hybrids. Since chloroplasts in Nicotiana are maternally inherited, these results demonstrate that polypeptides G and L are encoded by nuclear genes, are polymorphic variants of the PSII 23-kDa polypeptide, and are inherited in a Mendelian manner.Abbreviations kDa kilodalton - LS large subunit of Rubisco - M_r relative molecular mass - NEPHGE non-equilibrium pH gradient gel electrophoresis - PSII photosystem II - Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase - SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis - SS small subunit of Rubisco     

Biological productivity of Lake Towada,a north temperate,oligotrophic, kokanee fishery lake

During the Lake Towada survey from April through October 1998, we measured primary production at shore and offshore stations, and calculated crustacean zooplankton production from samples collected at the offshore station. We then analyzed these data and compared them with commercial fishery data in order to discuss the energy flow in this lake where kokanee (Oncorhynchus nerka) fisheries are one of the main businesses. At all stations, primary production was relatively high: 150–300mgCm^–2day^–1 in April–mid-June and lower at ca. 100mgCm^–2day^–1 thereafter. The variation in primary productivity could largely be explained by multiple regression models that included phytoplankton biomass and ambient nutrient conditions as independent variables. Among zooplankton, rotifers had their peak abundance in May, before the crustacean zooplankton (Daphnia longispina, Bosmina longirostris, and Acanthodiaptomus pacificus) population was well established. D. longispina dominated the crustacean zooplankton community in terms of biomass and production; their production during the study period made up 80% of crustacean community production (19.6gdry-wtm^–2), which was 40% of primary production during the survey. In July, when the abundance of D. longispina was particularly high, their daily production slightly exceeded daily primary production, which resulted in ca. 30% and 75% reduction in the amount of particulate organic carbon and chlorophyll a, respectively, during this period. The community ingestion rate of crustacean zooplankton, calculated from their daily production (D. longispina accounted for 90%) and the assumption that their gross production efficiency (K₁) was 60%, could roughly explain this reduction of particulate matter, corroborating previous studies that the grazing of D. longispina can significantly improve the water transparency of this lake. The catch of kokanee and pond smelt (Hypomesus transpacificus nipponensis, another important fish in the lake) during the survey corresponded to 1.1% of crustacean community production, and corresponded to 0.45% of the primary production, which is one of the highest recorded values. Bearing in mind that D. longispina was the major food item of planktivorous fish such as kokanee and pond smelt, the present study suggests that the energy transfer from phytoplankton to zooplankton to fish is outstandingly efficient, compared with other aquatic ecosystems, when D. longispina dominates in the lake.