Intermolecular protein interactions in solutions of calf lens alpha-crystallin. Results from 1/T1 nuclear magnetic relaxation dispersion profiles.

From analyses of the magnetic field dependence of 1/T1 (NMRD profiles) of water protons in solutions of calf lens alpha-crystallin at several concentrations, we find two regimes of solute behavior in both cortical and nuclear preparations. Below approximately 15% vol/vol protein concentration, the solute molecules appear as compact globular proteins of approximately 1,350 (cortical) and approximately 1,700 (nuclear) kD. At higher concentrations, the effective solute particle size increases, reversibly, as evidenced by the appearance of spectra-like 14N peaks in the NMRD profiles and a change in the field and temperature dependence of 1/T1. At these higher concentrations, the profiles are very similar to those of calf gamma II-crystallin, a crystallin that undergoes an analogous transition near approximately 15% protein (Koenig, S. H., C.F. Beaulieu, R. D. Brown III, and M. Spiller, 1990. Biophys. J. 57:461-469). By comparison with recent analyses of NMRD results for solutions of immobilized proteins as models for the transition from protein solutions to tissue (Koenig, S. H., and R. D. Brown III. 1991. Prog. NMR Spectr. 22:487-567), we argue that alpha-crystallin solute behaves as aggregates approximately greater than 50,000 kD as protein concentration is progressively increased above 15%. Finally, the concentration dependence of the NMRD profiles of alpha- and gamma II-crystallin can readily explain recent osmotic pressure data, in particular the intersection of the respective pressure curves at approximately 23% vol/vol (Vérétout, F., and A. Tardieu. 1989. Eur. Biophys. J. 17:61-68).     

Enhanced Antisense Effects Resulting from an Improved Streptolysin-O Protocol for Oligodeoxynucleotide Delivery into Human Leukaemia Cells

Abstract

An improved protocol for delivering oligodeoxynucleotides into the cytoplasm and nucleus of human cells in culture using streptolysin O is described. The new procedure permitted reduced target protein expression to result from antisense suppression of target mRNA levels.     

Ammonium ion-excreting cyanobacterial mutant as a source of nitrogen for growth of rice: A feasibility study

Summary Growth of rice plants in a nitrogen-free medium was enhanced by inoculation with a nitrogenase-derepressed mutant (strain SA-1) of a cyanobacterium,Anabaenavariabilis which excretes NH₄⁺ into the medium. Both total dry weight and nitrogen content of rice plants were substantially increased in the presence of the mutant strain but not with the wild type parent, strain SA-0.     

Ammonia-excreting mutant strain of the cyanobacterium Anabaena variabilis supports growth of wheat

Summary Growth of wheat in a nitrogen-free hydroponic co-culture with a mutant strain of the cyanobacterium Anabaena variabilitis (strain SA-1) was enhanced over plants grown with the parent strain SA-0. This increase was achieved in the dry weight, grain yield, and total nitrogen content of the plants. Nitrogenase activity of the mutant strain SA-1 was increased in a co-culture of the cyanobacterial mutant with wheat plants compared to the activity of the wild-type strain in association with wheat.Offprint requests to: M. Gunasekaran     

Fructose uptake and influence on growth of and nitrogen fixation by Anabaena variabilis

Fructose is specifically taken up by nitrogen-fixing cultures of Anabaena variabilis in the light and lowers the doubling time from 24 to 8 h. The kinetics for both fructose-dependent growth and fructose uptake are exponential. The apparent Km for fructose uptake in N2-fixing cultures is 160 microM for cells not previously exposed to fructose and 50 microM in cells adapted to fructose. Picomolar amounts of [14C]fructose are scavenged from the medium and accumulate in filaments. Heterocysts of fructose-adapted filaments accumulate 14C from fructose within 20 min. Short-term experiments with fructose-starved cultures provide evidence that nitrogenase activity, protein, and chlorophyll content change within one generation time upon addition of fructose. In long-term experiments, the amount of fructose initially present in the medium determines heterocyst number and packed-cell volume. Photosynthetic oxygen evolution and amounts of chlorophyll decrease with exogenous fructose concentrations greater than 20 mM.