Abstract:In this paper, TPK activity from human peripheral blood lymphocyte membrane was measured using synthetic PGAT as substrate. The kinase activity was more strongly inhibited by a synthetic compound,1-isopropyl-6-amino-thieno (3,4-d) pyrimidine-2, 4-dione (IATPD); IC50 was about 300μmol/L. Lineweaver-Burk plots revealed that IATPD was competitively with ATP and noncompetitively with PGAT to inhibit the kinase activity.
Abstract:A systematic investigation of the property of the SDS-protein complex and the influence of the detergent sodium doedecyl sulphate (SDS) on protein purification is reported. The data demonstrate that SDS in the sample doesn't disturb gel permeation and hydro xyapatite chromatographic separation, but reduces the separation efficiency and protein recovery in hydrophobic interaction, affinity and reversed-phased chromatographic methods. A successful separation of many membrane and inclusion body proteins in genetic engineering will therefore require new concepts in column design as well as critical revision of the choice of mobile phases. This study provides important data on membrane and inclusion body proteins purification.
Abstract:Sprague - Dawley rats were used to study the effects of insulin on theapo AⅠ, CⅡ and CⅢ gene expressions. Using 32P-labelled apo AⅠ, CⅡ and CⅢ recombinant DNA as probes, we determined the mRNA amounts of apo AⅠ, CⅡ and CⅢ in the liver and intestine of rat injected with insulin (1.5U/Kg weight of rat) by dot blot hybridization method. After injecting insulin, the mRNA amounts of apo AⅠ in liver and intestine were obviously decreased, to a lowest level (42%) at 6 h interval and restored to that of control group at 24h interval. Also, after injecting insulin, the mRNA amounts of apo CII in liver and intestine were declined remarkably, to a lowest level (57%) at 12h interval, still lower than that of the control group (79%) at 24h interval. But the mRNA amounts of apo CIII showed no significant changes after rejecting insulha both in liver and intestine when compared with the control group (P＞0.05). The experiment was repeated.12h after injecting insulin, the mRNA amounts of apo Al, CII and CIII in liver and apo A1 and CII in intestine were obviously decreased (45%, 51% and 54%, 35% and 37%, respectively, P＜0.01, n=5) and the mRNA amounts of apo CIII in intestine remained unchange (P＞0.05, n= 5). These resulls revealed that insulin could evidently affect the gene expression of apolipoproteins.
