Size exclusion chromatography

The diseaseFusarium head blight (scab) causes severe problems for farmers and for the industries that use cereals. It is likely that the fungi that cause scab (Fusariumspp.) use various enzymes when they invade grains. We are studying enzymes that the fungi may use to hydrolyze grain proteins. To do this,Fusarium culmorumwas grown in a gluten-con-taining medium from which an alkaline serine proteinase with a molecular mass of 28.7 kDa was puri®ed by size-exclusion and cation exchange chromatographies....

10p research12 01-06-2013 54 5   Download

a-Crystallin, a molecular chaperone and lens structural protein protects soluble enzymes against heat-induced aggregation and inactivation by a variety of molecules. In this study we investigated the chaperone function ofa-crys-tallininamorephysiological systeminwhicha-crystallinwas incorporated into red cell
ghosts. Its ability to protect the intrinsic membrane protein Na/K-ATPase from external stresses was studied. Red cell ghosts were created by lysing the red cells and removing cytoplasmic contents by size-exclusion chromatography. ...

7p fptmusic 16-04-2013 48 2   Download

Integrase (IN) is the retroviral enzyme responsible for the integration of the DNA copy of the retroviral genome into the host cellDNA. TheC-terminal domainof INis involved in DNA binding and enzyme multimerization. We previ-ously performed single amino acid substitutions in the C-terminal domain of the avian leukemia and sarcoma vir-uses (ALSV) IN [Moreauet al. (2002). Arch. Virol.147, 1761–1778].

13p fptmusic 12-04-2013 42 4   Download

The guanidinium chloride- and urea-induced unfolding of FprA, a mycobacterium NADPH-ferredoxin reductase, was examined in detail using multiple spectroscopic techniques, enzyme activity measurements and size exclusion chromatography. The equilibrium unfolding of FprA by urea is a cooperative process where no stabilization of any partially folded inter-mediate of protein is observed. In comparison, the unfolding of FprA by guanidinium chloride proceeds through intermediates that are stabilized by interaction of protein with guanidinium chloride. ...

0p awards 05-04-2013 36 3   Download

In an attempt to isolate lipopolysaccharide fromSpirocha-eta aurantia, Darveau-Hancock extraction of the cell mass was performed.Whileno lipopolysaccharidewas found, two carbohydrate-containing compounds were detected. They were resolved by size-exclusion chromatography into high molecularmass(LGLA) and low molecular mass (LGLB) fractions. Here we present the results of the analysis of the glycolipid LGLB.

11p awards 05-04-2013 34 2   Download

A hallmark ofa-crystallin-type small heat shock proteins (sHsps) is their highly dynamic oligomeric structure which promotes intermolecular interactions involved in subunit exchange and substrate binding (chaperone-like activity). We studied the oligomeric features of two classes of bacterial sHsps by size exclusion chromatography and nanoelectro-spray mass spectrometry.Proteins of both classes formed large complexes that rapidly dissociated upon dilution and at physiologically relevant heat shock temperatures....

10p dell39 03-04-2013 45 3   Download

Human heat shock protein of apparent molecular mass 20 kDa (Hsp20) and its mutant, S16D, mimicking phos-phorylation by cyclic nucleotide-dependent protein kinases, were cloned and expressed inEscherichia coli. The proteins were obtained in a homogeneous state without utilization of urea or detergents. On size exclusion chromatography at neutral pH, Hsp20 and its S16D mutant were eluted as symmetrical peaks with an apparent molecular mass of 55–60 kDa.

12p dell39 03-04-2013 46 3   Download

Chicken pancreatic phospholipase A2 (ChPLA2 ) was purified from delipi-dated pancreases using ammonium sulfate and ethanol precipitation, followed by sequential column chromatography steps on MonoQ Sepha-rose and size exclusion HPLC columns. ChPLA2 was found to be a non-glycosylated monomeric protein with a molecular mass of 14 kDa and a specific activity of 400 UÆmg )1 in the presence of 1 mmsodium taur-odeoxycholate and 4 mmCaCl2with phosphatidylcholine as substrate.

10p vinaphone15 25-02-2013 38 4   Download

In a biological environment, nanoparticles immediately become covered by an evolving corona of biomolecules, which gives a biological identity to the nanoparticle and determines its biological impact and fate. Previous efforts at describing the corona have concerned only its protein content. Here, for the first time, we show, using size exclusion chromatography, NMR, and pull-down experiments, that copolymer nanoparticles bind cholesterol, tri-glycerides and phospholipids from human plasma, and that the binding reaches saturation. ...

10p viettel02 22-02-2013 37 4   Download