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Biomedical subjects

R A Cooper

Publications and source records attributed to R A Cooper.

At least 307 records · Page 17Linked to original sources

The purification and properties of Escherichia coli methylglyoxal synthase.

1. Methylglyoxal synthase was purified over 1500-fold from glycerol-grown Escherichia coli K 12 strain CA 244. The purified enzyme was inactivated by heat or proteolysis, had a molecular weight of approx. 67000, a pH optimum of 7.5 and was specific for dihydroxyacetone phosphate with K(m) 0.47mm. 2. The possibility that a Schiff-base intermediate was involved in the reaction mechanism was investigated but not confirmed. 3. The purified enzyme lost activity, especially at low temperature, but could be stabilized by P(i). Two binding sites for P(i) may be present on the enzyme. Of other compounds tested only the substrate, dihydroxyacetone phosphate, and bovine serum albumin showed any significant stabilizing effect. 4. Phosphoenolpyruvate, 3-phosphoglycerate, PP(i) and P(i) were potent inhibitors of the enzyme. Kinetic experiments showed that PP(i) was apparently a simple competitive inhibitor, but inhibition by the other compounds was more complex. In the presence of P(i) the enzyme behaved co-operatively, with at least three binding sites for dihydroxyacetone phosphate. 5. It is proposed that methylglyoxal synthase and glyceraldehyde 3-phosphate dehydrogenase play important roles in the catabolism of the triose phosphates in E. coli. Channelling of dihydroxyacetone phosphate via methylglyoxal would not be linked to ATP formation and could be involved in the uncoupling of catabolism and anabolism.

Acetone↗

Loss of membrane components in the pathogenesis of antibody-induced spherocytosis.

The infusion of hyperimmune agglutinating antibodies into man or animals causes spherocytosis and hemolysis. The mechanism of spherocytosis was studied in rats given rabbit anti-rat red cell antiserum intravenously. During the 18 hr after antibody infusion, a time before the onset of reticulocytosis, hematocrits fell from 40.6 to 27.6%. However, no change occurred in mean cell volume, mean cell hemoglobin content, or the red cell concentrations of potassium or adenosine triphosphate (ATP). There was a progressive loss of membrane constituents and membrane surface area which followed first order kinetics. At 18 hr membrane cholesterol had decreased 23.5%, phospholipid 26.3%, protein 4.7%, and surface area (calculated from a measure of osmotic fragility) 14.2%. There was no change in the per cent composition of the various phospholipids. Similar changes occurred in animals splenectomized before receiving antibody.These studies demonstrate that spherocytosis induced by heterologous agglutinating antibodies in vivo results from a loss of surface area with no accompanying change in cell volume or in the concentration of the major intracellular constituents. It is caused by a process acting at the cell surface leading to the loss of lipid-rich, protein-poor components of the red cell membrane.

Adenosine Triphosphate↗

An analysis of lipoproteins, bile acids, and red cell membranes associated with target cells and spur cells in patients with liver disease.

Most patients with stable cirrhosis of the alcoholic have "target" red cells; however, a minority have "spur" cells and severe hemolytic anemia. These two syndromes were studied in 27 patients with target cells and 17 patients with spur cells, all of whom had advanced cirrhosis. The cholesterol and phospholipid content of red cell membranes effectively distinguished target cells from spur cells. Target cells alone were rich in lecithin, and both the cholesterol/phospholipid and cholesterol/lecithin mole ratios were greater in spur cells. The cholesterol/phospholipid mole ratio of both types of red cells correlated closely with the free cholesterol saturation of serum lipoproteins, as defined by the amount of free cholesterol relative to phospholipid and protein in these lipoproteins. Lecithin: cholesterol acyltransferase (LCAT) activity was decreased in most patients with target cells and spur cells; however, the relationship between this activity and the lipid abnormalities observed was weak. Serum bile acid levels also correlated poorly with serum and cell lipids. However, in patients with target cells the amount of cholic and deoxycholic acids in serum was approximately equal to the amount of chenodeoxycholic acid, whereas in patients with spur cells chenodeoxycholic acid (the precursor of lithocholic acid) predominated.

Acetyltransferases↗

Migrations and growth of deep-sea lobsters, Homarus americanus.

In distinct contrast to the restricted movements of coastal stocks of lobsters (Homarus americanus), those inhabiting the outer continental shelf undertake extensive seasonal migrations. Of 5710 tagged lobsters released on the outer continental shelf off New England from April 1968 to June 1969, 400 had been recaptured by April 1970. The distribution of the recoveries demonstrated shoalward migration in spring and summer and a return to the edge of the shelf in fall and winter. Deep-sea lobsters have a faster rate of growth than coastal lobsters; growth increments at molting and the frequency of molting are greater.

Animals↗