Report on scientific research

The broad objective of research reported here was to develop a multi-publishing workflow for student atlases.

162p runthenight05 01-03-2023 5 3   Download

The dinitrogenase component proteins of the conventional Mo nitrogenase (MoFe protein) and of the alternative Fe-only nitrogenase (FeFe protein) were both isolated and purified from Rhodobacter capsulatus, redox-titrated according to the same procedures and subjected to an EPR spectroscopic comparison. In the course of an oxidative titration of the MoFe protein (Rc1Mo) three significant S ¼ 1/2 EPR signals deriving from oxidized states of the P-cluster were detected: (1) a rhombic signal (g ¼ 2.07, 1.96 and 1.

12p system191 01-06-2013 31 4   Download

DNA of prokaryotes is in a nonequilibrium structural state, characterized as ÔactiveÕ DNA supercoiling. Alterations in this state a€ect many life processes and a homeostatic control of DNA supercoiling has been suggested [Menzel, R. & Gellert, M. (1983) Cell 34, 105±113]. We here report on a new method for quantifying homeostatic control of the high-energy state of in vivo DNA. The method involves making small perturbation in the expression of topoisomerase I, and measuring the e€ect on DNA supercoiling of a reporter plasmid and on the expression of DNA gyrase....

8p system191 01-06-2013 34 5   Download

The human b-globin gene locus is the subject of intense study, and over the past two decades a wealth of information has accumulated on how tissue-specific and stage-specific expression of its genes is achieved. The data are extensive and it would be difficult, if not impossible, to formulate a comprehensive model integrating every aspect of what is currently known. In this review, we introduce the fundamental characteristics of globin locus regulation as well as questions on which much of the current research is predicated.

11p system191 01-06-2013 47 4   Download

The importance of two putative Zn2+-binding (Asp347, Glu429) and two catalytic (Arg431, Lys354) residues in the tomato leucine aminopeptidase (LAP-A) function was tested. The impact of substitutions at these positions, corresponding to the bovine LAP residues Asp255, Glu334, Arg336, and Lys262, was evaluated in His6–LAP-A fusion proteins expressed in Escherichia coli. Sixty-five percent of the mutant His6–LAP-A proteins were unstable or had complete or partial defects in hexamer assembly or stability. The activity of hexameric His6–LAP-As on Xaa-Leu and Leu-Xaa dipeptides was tested.

11p system191 01-06-2013 40 4   Download

Bacteroides vulgatus has been shown to be involved in the aggravation of colitis. Previously, we separated two potent virulence factors, capsular polysaccharide (CPS) and lipopolysaccharide (LPS), from a clinical isolate of B. vulgatus and characterized the structure of CPS. In this study, we elucidated the structures of O-antigen polysaccharide (OPS) and lipid A in the LPS. LPS was subjected to weak acid hydrolysis to produce the lipid A fraction and polysaccharide fraction.

7p system191 01-06-2013 50 8   Download

Guanosine triphosphate nucleotide analogues such as GppNHp (also named GMPPNP) or GTPcS are widely used to stabilize rapidly hydrolyzing protein-nucleotide complexes and to investigate biochemical reaction pathways. Here we describe the chemical synthesis of guanosine 5¢-O-(c-amidotriphosphate) (GTPcNH2) and a new synthesis of guanosine 5¢-O-(c-fluorotriphosphate) (GTPcF). The two nucleotides were characterized using NMR spectroscopy and isothermal titration calorimetry. Chemical shift data on 31P, 19F and 1H NMR resonances are tabulated.

9p system191 01-06-2013 32 5   Download

Many groups of proteins play important roles in the cell’s response to various stresses. The molecular chaperone GroEL of Escherichia coli represents one such highly conserved family of stress proteins. We have observed that isolated GroEL complexes from stationary cultures contain various polypeptides that can be released from the chaperonin by GroES and/or ATP, and identified two such polypeptides as the proteins GatY and UP12.

9p system191 01-06-2013 28 3   Download

Agrobacterium tumefaciens is a Gram-negative, phytopathogenic bacterium and is characterized by an unique mode of action on dicotyledonous plants: it is able to genetically modify the host, and because of this feature, it is used as a tool for transgenic plants. Many experiments have demonstrated that lipopolysaccharides (LPSs) play an important role for the disease development, as they are involved in the adhesion process of the bacterium on the plant cell wall.

4p system191 01-06-2013 32 4   Download

Chromatin is a highly dynamic structure that plays an essential role in regulating all nuclear processes that utilize the DNA template including DNA repair, replication, transcription and recombination. Thus, the mechanisms by which chromatin structures are assembled and modified are questions of broad interest. This minireview will focus on two groups of proteins: (a) histone chaperones and (b) ATPdependent chromatin remodeling machines, that co-operate to assemble DNA and histone proteins into chromatin....

7p research12 01-06-2013 32 4   Download

By replacing specific amino acids at positions 112, 147 and 152 of the human aldosterone synthase (CYP11B2) with the corresponding residues from human, mouse or rat 11b-hydroxylase (CYP11B1), we have been able to investigate whether these residues belong to structural determinants of individual enzymatic activities. When incubated with 11-deoxycorticosterone (DOC), the 11b-hydroxylation activity of the mutants was most effectively increased by combining D147E and I112P (sixfold increase).

10p research12 01-06-2013 36 4   Download

Insulin is the principal regulatory hormone involved in the tight regulation of fuel metabolism. In response to blood glucose levels, it is secreted by the b cells of the pancreas and exerts its effects by binding to cell surface receptors that are present on virtually all cell types and tissues. In humans, perturbations in insulin function and/or secretion lead to diabetes mellitus, a severe disorder primarily characterized by an inability to maintain blood glucose homeostasis. Furthermore, it is estimated that 90–95% of diabetic patients exhibit resistance to insulin action....

10p research12 01-06-2013 34 4   Download

The structure of post-translational modifications of human heparin cofactor II isolated from human serum and from recombinant Chinese hamster ovary cells and their effects on heparin binding have been characterized. Oligosaccharide chains were found attached to all three potential N-glycosylation sites in both protein preparations. The carbohydrate structures of heparin cofactor II circulating in blood are complex-type diantennary and triantennary chains in a ratio of 6 : 1 with the galactose being 90% sialylated with a2 fi 6 linked N-acetylneuraminic acid....

12p research12 01-06-2013 44 3   Download

Csk-binding protein⁄phosphoprotein associated with glycosphingolipid-enriched domains is a transmembrane adaptor protein primarily involved in negative regulation of T-cell activation by recruitment of C-terminal Src kinase (Csk), a protein tyrosine kinase which represses Src kinase activity through C-terminal phosphorylation. Recruitment of Csk occurs via SH2-domain binding to PAG pTyr317, thus, the interaction is highly dependent on phosphorylation performed by the Src family kinase Fyn, which docks onto PAG using a dual-domain binding mode involving both SH3- and SH2-domains of Fyn....

12p research12 29-04-2013 40 3   Download

The binding of lipopolysaccharide (LPS, also known as bacterial endotoxin) to human hemoglobin is known to result in oxidation of hemoglobin to methemoglobin and hemichrome. We have investigated the effects of the LPSs from smooth and rough Escherichia coliandSalmonella minnesotaon the rate of oxidation of native oxyhemoglobin A0and hemoglobin cross-linked between thea-99 lysines.

6p research12 23-04-2013 44 3   Download

The effects of pressure on structure and activity of recom-binant human acetylcholinesterase (rHuAChE) were inves-tigatedup toapressureof 300 MPausinggel electrophoresis under elevated hydrostatic pressure, fluorescence of bound 8-anilinonaphthalene-1-sulfonate (ANS) and activity meas-urements followingexposure tohighpressure.

11p research12 23-04-2013 36 1   Download

Our laboratory showed that bikunin, aKunitz-type protease inhibitor, suppresses 4b-phorbol 12-myristate 13-acetate (PMA)- or tumor necrosis factor-alpha (TNFa)-induced urokinase-type plasminogen activator (uPA) expression in different cell types. In addition to its effects on protease inhibition, bikunincouldbemodulatingother cellular events associated with the metastatic cascade.

13p research12 23-04-2013 42 2   Download

Palytoxin stimulated a cation-dependent short-circuit cur-rent (Isc) in rat distal and proximal colon in a concen-tration-dependent fashion when applied to the mucosal surface of the tissue. The distal colon exhibited a higher sensitivity to the toxin. The palytoxin-induced Isc was blocked by vanadate but was resistant to ouabain or scilliroside, suggesting the conversion of a vanadate-sen-sitive H + /K + -ATPase into an electrogenic cation trans-porter.

7p research12 23-04-2013 58 3   Download

The thermal unfoldingofmyosinsubfragment 1 (S1) cleaved by trypsin was studied by differential scanning calorimetry. In the absence of nucleotides, trypsin splits the S1 heavy chain into three fragments (25, 50, and 20 kDa). This cleavage has no appreciable influence on the thermal unfolding of S1 examined in the presence of ADP, in the ternary complexes of S1 with ADP and phosphate analogs, such as orthovanadate (Vi) or berylliumfluoride (BeFx), and in the presence of F-actin.

11p tumor12 22-04-2013 34 3   Download

The presence of extraN- andC- terminal residues can play a major role in the stability, solubility and yield of recombi-nant proteins. Pfg27 is a 27Ksoluble protein that is essential for sexual development in Plasmodium falciparum.It was over-expressed using the pMAL-p2 vector as a fusion pro-teinwith themaltosebindingprotein. Sixdifferent constructs were made and each of the fusion proteins were expressed and purified.

5p tumor12 22-04-2013 35 2   Download