Polyacrylamide gel electrophoresis (PAGE) provides a versatile, gentle, high resolution method for fractionation and physical-chemical characterization of molecules on the basis of size, conformation, and net charge. The polymerization reaction can be rigorously controlled to provide uniform gels of reproducible, measurable pore size over a wide range. This makes it possible to obtain reproducible relative mobility (Rf) values as physical-chemical constants. Application and extension of Ogston9s (random fiber) model for a gel allows for calculation of molecular volume, surface area, or radius, free mobility, and valence from RJ measurements at several gel concentrations, to calculate gel concentration for optimal resolution, and to predict behavior of macromolecules on gel gradients by computerized methods. Extension of classical moving boundary theory has been used to generate multiphasic buffer systems (providing selective stacking, unstacking, restacking, and preparative steady-state-stacking) with known operating characteristics for any pH at 0° and 25°C. A general strategy for isolation of macromolecules and for macromolecular mapping has been developed. Preparative scale PAGE is operational for milligram loads and feasible for gram quantities.

https://science.sciencemag.org/content/sci/172/3982/440.full.pdf

Polyacrylamide Gel Electrophoresis

Assembly of the mitochondrial membrane system. IV. Role of mitochondrial and cytoplasmic protein synthesis in the biosynthesis of the rutamycin-sensitive adenosine triphosphatase.

Glutamate plays a central role in a wide range of metabolic processes in bacterial cells. This review focuses on the involvement of glutamate in bacterial stress responses. In particular, it reviews the role of glutamate metabolism in response against acid stress and other stresses. The glutamate decarboxylase (GAD) system has been implicated in acid tolerance in several bacterial genera. This system facilitates intracellular pH homoeostasis by consuming protons in a decarboxylation reaction that produces γ-aminobutyrate (GABA) from glutamate. An antiporter system is usually present to couple the uptake of glutamate to the efflux of GABA. Recent insights into the functioning of this system will be discussed. Finally, the intracellular fate of GABA will also be discussed. Many bacteria are capable of metabolizing GABA to succinate via the GABA shunt pathway. The role and regulation of this pathway will be addressed in the review.

/pdf/role-of-glutamate-metabolism-in-bacterial-responses-towards-1gthwt1csq.pdf

Role of glutamate metabolism in bacterial responses towards acid and other stresses

http://www.jbc.org/content/250/14/5688.full.pdf

Enhanced differential synthesis of proteins in a mammalian cell-free system by addition of polyamines.

(1980). Ultrastructure, Physiology, and Biochemistry of Bacillus Thuringiensis. CRC Critical Reviews in Microbiology: Vol. 8, No. 2, pp. 147-204.

Ultrastructure, Physiology, and Biochemistry of Bacillus Thuringiensis

A cell-free system from Bacillus thuringiensis was found to actively incorporate phenylalanine into hot trichloroacetic acid-precipitable material in the presence of synthetic polynucleotide, ribosomes, S-100 supernatant, an energy-generating system, and guanosine triphosphate Phenylalanine incorporation was absolutely dependent on the presence of spermidine in addition to magnesium ions, even when highly purified ribosomes were used The spermidine effect could not be attributed to inhibition of nucleases The ribosomal and supernatant fractions from Escherichia coli and B thuringiensis could be substituted for each other in this system The spermidine requirement was shown to be limited to the B thuringiensis ribosome fraction

Spermidine Requirement for Bacillus thuringiensis Ribosomes in Cell-Free Phenylalanine Incorporation

Enzymatic analyses of Bacillus thuringiensis extracts suggested that a modified Krebs tricarboxylic acid cycle (without α-ketoglutarate dehydrogenase) can operate during sporulation in conjunction with the glyoxylic acid cycle and the γ-aminobutyric acid pathway.

John N. Aronson

Papers

Spermidine Requirement for Bacillus thuringiensis Ribosomes in Cell-Free Phenylalanine Incorporation

γ-Aminobutyric Acid Pathway and Modified Tricarboxylic Acid Cycle Activity During Growth and Sporulation of Bacillus thuringiensis

Square acrylamide gel columns and a novel gel slicer

Palo Verde (Parkinsonia aculeata L.) seed aminoacylase.

Small polypeptide components of the Bacillus thuringiensis parasporal crystalline inclusion.