Studies on the uptake of exogenous phosphoglycerides by Escherichia coli B cells

The uptake of exogenous lipids by Escherichia coli B was studied under various conditions. Lysophosphoglycerides were absorbed more readily than diacyl analogues when sonicated dispersions of a single lipid were used. When these same lipids were admixed with coliform lipids and dispersed as vesicles, the uptake of lysophosphoglyceride diminished and was approximately equal to that of diacyl analogues. Neutral lipids such as diglyceride, fatty acid, and cholesterol were also absorbed when admixed and dispersed as vesicles with coliform lipids. The uptake of lysophosphoglyceride was stimulated slightly by all divalent cations tested except Mg2+; monovalent cations were ineffective. Uptake was accompanied by conversion of lysophosphoglyceride to diacyl analogues. In the presence of Ca2+, lysophosphatidylethanolamine also formed a more polar lipid product yet unidentified. The uptake of lipid did not cause release of 3H label into the medium from cells that had been grown in [3H]acetate-containing medium. Also, the [3H]phosphoglyceride content of such labelled cells remained constant. Thus the uptake process did not involve exchange of membrane lipid with the medium or an enhanced hydrolysis of endogenous lipids, but it represented a net gain of lipid by the cell. The uptake did not seem to involve a stable adsorption of lipid at the surface of the cell as could be judged from electron microscopic examination of the cells after incubation; the cell surfaces were devoid of adsorbed vesicle or liposomal types of structures and did not display evidence of expansion. [3H]Cholesterol-[32P]phospholipid mixtures were taken up without a change in isotopic ratio. This result together with the other evidence presented indicate a net uptake of exogenous lipid by a process likely involving fusion.     

Mitochondrial genome diversity in soybean: repeats and rearrangements

Mitochondrial (mt) genome organization in soybean was examined at the molecular level. This study builds upon previous reports that four soybean cytoplasmic groups, Bedford, Arksoy, Lincoln, and soja-forage, are differentiated by polymorphisms detected with a 2.3 kb Hind III mtDNA probe [12]. The variation detected results from DNA alterations in a region within and around a 4.8 kb repeat. The Bedford-type cytoplasm is the only cytoplasm that contains copies of a 4.8 kb repeat in four different genomic environments, evidence that it is recombinationally active. The Lincoln- and Arksoy-type cytoplasms each contain two copies of the repeat, as well as unique fragments that appear to result from rare recombination events outside, but near, the repeat. The soja-forage-type cytoplasm contains no complete copies of the repeat, but does contain a unique truncated version of the repeat. Sequence analysis indicates that the truncation is a result of recombination across a 9 bp repeated sequence, CCCCTCCCC. The structural rearrangements that have occurred in the region surrounding the 4.8 kb repeat may provide a means to dissect species relationships and evolution within the subgenus soja.     

Isolation of subpopulations of murine epidermal cells using monoclonal antibodies against differentiation-related cell surface molecules

Abstract. Monoclonal antibodies (mAbs) against epithelial cells were prepared by immunization of rats with lyophilized murine epithelia. Screening against tissue sections and epithelial cell suspensions permitted identification of mAbs against surface molecules that are expressed early in cell differentiation. Staining with These mAbs followed by fluorescence-activated cell sorting enabled isolation of subpopulations of basal epithelial cells. Staining these subpopulations with antibodies against known differentiation markers (cytokeratins and bullous pemphigoid antigen) and measurements of cell size indicated that they represented fractions of the basal cell population in sequential stages of early differentiation. Labeling mice with bromodeoxyuridine at various limes prior to cell isolation showed that the least-differentiated basal cells cycle more slowly than those at later stages, data which support the concept of a differentiation-related, hierarchical pattern of organization of the proliferative compartment.     

Kinetic mechanism of formininotransferase from porcine liver.

Formiminotransferase (EC 2.1.2.5) and cyclodeaminase (EC 4.3.1.4) constitute an enzyme complex that catalyses two sequential metabolic reactions. The activity of native formiminotransferase can be measured without interference from cyclodeaminase, and its kinetic mechanism has been investigated. Although initial velocity plots yield families of parallel lines suggesting that the transferase utilizes a ping-pong mechanism, product inhibition and alternate substrate studies with tetrahydropteroic acid clearly show the mechanism to be sequential. Of the possible mechanisms compatible with these observations, several could be ruled out through the effects of various dead-end inhibitors. The data indicate that the transferase mechanism is rapid equilibrium random with formation of a dead-end complex enzyme-tetrahydrofolate-glutamate.