Phosphorylation of rat kidney pyruvate kinase type L by cyclic 3',5'-AMP-dependent protein kinase.

Pyruvate kinase (ATP:pyruvate 2-O-phosphotransferase, EC 2.7.1.40) type L was partly purified from rat kidney. During the last two purification steps, the incorporation of [32P]phosphate into protein on incubation with [32P]ATP and cyclic 3',5'-AMP-dependent protein kinase was found to parallel the pyruvate kinase activity. After phosphorylation of the enzyme, a major radioactive band with a molecular weight of 57 000 was found on polyacrylamide gel electrophoresis [32P]Phosphorylserine was isolated from the kidney pyruvate kinase. Immunological identity was found between the liver and kidney pyruvate kinases type L. By autoradiography of high-voltage electropherograms after partial acid hydrolysis of the phosphorylated rat liver and kidney pyruvate kinases type L, identical results were obtained. The affinity for phosphoenolpyruvate was found to be decreased by phosphorylation of the enzyme with a change in the apparent Km from 0.15 mM to 0.35 mM. After incubation of the phosphorylated kidney pyruvate kinase with phosphatase the phosphoenolpyruvate saturation curve was found to be identical to that for the unphosphorylated enzyme. Thus, the activity of the rat kidney pyruvate kinase type L is with all probability regulated by a reversible phosphorylation-dephosphorylation reaction, thereby indicating that hormonal regulation of gluconeogenesis via cyclic AMP may be of importance in the renal cortex.     

Investigation of immunosuppressive properties of inactivated human immunodeficiency virus and possible neutralization of this effect by some patient sera

Retroviral infections are accompanied by immunosuppression in a variety of species. For feline leukemia virus, the immunosuppression has been ascribed to the transmembrane envelope protein, p15E, which suppresses the proliferative responses of cat, mouse, and human lymphocytes. A similar suppressive effect has been shown for a lysate of human immunodeficiency virus (HIV), strain HTLV-IIIB. Here we determined that detergent-disrupted HTLV-IIIB lystate exerted a strong suppressive effect on PHA-stimulated lymphocytes. Preparations of whole virions, a lysate of a local HIV isolate grown on MP-6 cells, and a commercially obtained UV and psoralene-inactivated lysate were examined and demonstrated to have a similar suppressive effect. The HIV lysate was not directly cytotoxic to lymphocytes and did not contain tumor necrosis factor or lymphotoxin. The HIV lysate specifically suppressed the proliferation of a range of hemopoietic cell lines from man and mouse including three EBV transformed CD4- and IL-2 receptor-negative B-cell lines. The lysate also suppressed the formation of human bone marrow colonies, whereas the lysate had only a slight or no effect on fibroblasts. The suppression of lymphocyte proliferation was not abrogated by addition of IL-2 or IL-1 and the HIV lysate inhibited the expression of IL-2 receptors on suboptimal PHA-stimulated mononuclear cells. The suppressive factor(s) has not been characterized in molecular terms, but suppressive activity was recovered in fractions with a molecular weight of about 67,000 and in both the glycoprotein fraction and in the glycoprotein-depleted fraction of the HIV lysate. Sera from one-third of a small series (N = 13) of individuals with antibodies to HIV seem to be able to neutralize the suppressive properties of HIV lysate in cultures.     

Hydrocarbon uptake in hydrocarbon fermentations

Candida lipolytica (strain ATCC 8662) was grown on a simple defined medium with n-hexadecane as the main carbon Source under batch fermentation conditions. The relative importance of the cells growing in the aqueous phase on the overall kinetics was studied. The effect of interfacial tension, unoccupied interfacial area, and pseudosolubility on the specific growth was also studied. Results are presented and discussed here.     

Significance of low-temperature growth associated with the fecal coliform response, indole production, and pectin liquefaction in Klebsiella

In the genus Klebsiella, the growth respnse in nutient broth at 10 degrees C correlates inversely with the operational definition of a fecal coliform and not merely with the ability to grow at 44.5 degrees C. Of the fecal coliform-positive Klebsiella, 97% did not grow at 10 degrees C after 72 h of incubation. Conversely, 97% of the fecal coliform-negative isolates grew at 10 degrees C. The amount of growth at 10 degrees C varied among the fecal coliform-negative isolates and was found to correlate with indole production and pectin liquefaction. Low-temperature growth associated with specific biochemical tests can be used to differentiate several groups in the genus Klebsiella. Three main groups were discerned. Group I consists of indole-negative, pectin-nonliquefying, fecal coliform-positive isolates that do not grow at 10 degrees C. Group II isolates are differentiated from group I by a fecal-coliform-negative response and growth at 10 degrees C. Group III are indole-positive, pectin-liquefying, fecal coliform-negative isolates that grow at 10 degrees C. In our culture collection, isolates of group I are most frequently of human/animal clinical origins, whereas isolates of groups II and III are predominantly derived from the environment.