Cytoplasmic extracts prepared from Xenopus laevis eggs are used for the reconstitution of a wide range of processes in cell biology, and offer a unique environment in which to investigate the role of cytoplasmic mechanics without the complication of preorganized cellular structures. As a step toward understanding the mechanical properties of this system, we have characterized the rheology of cr...

The characteristics of glucose transport were investigated in erythrocytes of a primitive vertebrate, the Pacific hagfish (Eptatretus stouti) Lockington. Transport of glucose by intact hagfish erythrocytes and by phospholipid vesicles reconstituted with noctylglucoside extract of hagfish erythrocyte membranes was rapid and mediated by a saturable stereospecific mechanism sensitive to inhibition...

A low molecular weight cytoplasmic protein (Mr 19,000-22,000) has been reported to be overexpressed in some multidrug-resistant cells. We have found that a cytoplasmic protein with a molecular weight of 22,000 is highly expressed in the human myelogenous leukemia K562 cells resistant to Adriamycin (K562/ADM). The Mr 22,000 protein was shown to be one of the major calcium-binding proteins in the...

The aim of this work was to establish, on a quantitative basis, the subcellular distribution of the enzyme system that converts arachidonic acid into prostaglandin (PG) E2 in mouse resident peritoneal (MRP) macrophages. Kinetic studies were conducted on cell-free extracts derived from cells cultivated for 1 d, using [1-14C]arachidonic acid as substrate and measuring the label in PGE2 after extr...

Intact Rickettsia prowazekii was radiolabeled with the glucose oxidase-lactoperoxidase method of iodination. Separation of the rickettsial extract into cytoplasmic, outer and inner membrane fractions demonstrated that the outer membrane was preferentially labeled. Analysis of the polypeptides of these fractions on high-resolution slab polyacrylamide gels showed that most of the 125I was in poly...

We have investigated whether poly(C)-binding protein (PCBP)-1 and PCBP-2 interact with the hepatitis C virus (HCV) 5' untranslated region. Our results demonstrate that glutathione S-transferase (GST)-PCBP-1 and GST-PCBP-2 fusion proteins bind specifically to the HCV 5' untranslated region. An antiserum raised against PCBP-2 induced a supershift after incubation with RNA-protein complexes formed...

We have adapted a method to map cell surface proteins and to monitor the effect of specific regulatory RNAs on the surface composition of the bacteria. This method involves direct labeling of surface proteins of living bacteria using fluorescent dyes and a subsequent separation of the crude extract by 2D gel electrophoresis. The strategy yields a substantial enrichment in surface proteins over ...

The in vitro antimicrobial activity of different fractions obtained from rhizome of Curcuma longa was investigated against standard strain and clinical isolates of Staphylococcus aureus. The clinical isolates were found more sensitive for different fractions, than the standard strain of S. aureus. Scanning electron microscopic observations revealed that test pathogen treated with C. longa extra...

A two-dimensional gel electrophoresis (2-DE)-based proteomic approach was used to study a transgenic mouse model of acerbated dilated cardiomyopathy in which the small monomeric GTPase, Rac1, was constitutively expressed exclusively in the myocardium. A subfractionation procedure allowed for the focused analysis of both cytoplasmic and myofilament protein-enriched extracts of ventricular tissue...

Cell suspension cultures of chickpea (Cicer arietinum L.) were used to prepare protoplasts and vacuoles. The vacuolar preparation revealed only slight contaminations of cytoplasmic marker enzymes. HPLC analysis of the vacuolar extract showed that the malonylglucosides of isoflavones, isoflavanones and pterocarpans are exclusively located in the vacuole. Experi­ ments designed to determine the s...