Protein precipitation

This study aimed to develop and validate the high performance liquid chromatography (HPLC-DAD) method combined with liquid-liquid extraction followed by an additional clean-up for the simultaneous determination of TMP and SMX in human plasma.

8p visarada 28-04-2025 1 1   Download

The three genespduCDEencoding the diol dehydratase of Lactobacillus collinoides, have been cloned for overexpres-sion in the pQE30 vector. Although the three subunits of the protein were highly induced, no activity was detected in cell extracts. The enzyme was therefore purified to near homo-geneity by ammoniumsulfate precipitation and gel filtration chromatography. In fractions showing diol dehydratase activity, three main bands were present after SDS/PAGE with molecular masses of 63, 28 and 22 kDa, respectively....

7p tumor12 22-04-2013 52 2   Download

A cellulase (endo-b-1,4-glucanase, EC 3.2.1.4) was purified from the gut of larvae of the yellow-spotted longicorn beetle Psacothea hilarisby acetone precipitation and elution from gels after native PAGE and SDS/PAGE with activity staining. The purified protein formed a single band, and the molecular mass was estimated to be 47 kDa. The purified cellulasedegradedcarboxymethylcellulose (CMC), insoluble cello-oligosaccharide (average degree of polymerization 34) and soluble cello-oligosaccharides longer than cellotriose, but not crystalline cellulose or cellobiose. ...

6p tumor12 20-04-2013 46 5   Download

The peptide hormone endothelin transmits various signals throughGprotein-coupled receptors, the endothelin type A (ETAR) and B (ETBR) receptors. Caveolae are specialized lipid rafts containing polymerized caveolins. We examined the interaction of ETBR with caveolin-1, expressed in Sf9, COS-1, and HEK293 cells, and its effects on the subcellular distribution and the signal transduction of ETBR.

12p fptmusic 16-04-2013 28 2   Download

Stringent factor is a ribosome-dependent ATP:GTP pyrophosphoryl trans-ferase that synthesizes (p)ppGpp upon nutrient deprivation. It is activated by unacylated tRNA in the ribosomal amino-acyl site (A-site) but it is unclear how activation occurs. A His-tagged stringent factor was isolated by affinity-chromatography and precipitation. This procedure yielded a protein of high purity that displayed (a) a low endogenous pyrophosphoryl transferase activity that was inhibited by the antibiotic tetracycline; ...

0p awards 05-04-2013 47 2   Download

Cell-free expression has become a highly promising tool for the fast and efficient production of integral membrane proteins. The proteins can be produced as precipitates that solubilize in mild detergents usually without any prior denaturation steps. Alternatively, membrane proteins can be syn-thesized in a soluble form by adding detergents to the cell-free system.

15p dell39 27-03-2013 52 4   Download

After the successful cloning of the first gene for a polyglutamine disease in 1991, the expanded polyglutamine tract in the nine polyglutamine disease proteins became an obvious therapeutic target. Early hypotheses were that misfolded, precipitated protein could be a universal pathogenic mechanism. However, new data are accumulating on Huntington’s disease and other polyglutamine diseases that appear to contradict the toxic aggregate hypothesis.

11p galaxyss3 07-03-2013 56 5   Download

Small heat shock proteins (sHsps) are intracellular molecular chaperones that prevent the aggregation and precipitation of partially folded and destabilized proteins. sHsps comprise an evolutionarily conserved region of 80–100 amino acids, denoted thea-crystallin domain, which is flanked by regions of variable sequence and length: the N-terminal domain and the C-terminal extension.

14p vinaphone15 28-02-2013 38 2   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 42 4   Download

To elucidate the architectural principle of protein structure, we focused on sequestration from solvent, which is a common characteristic of folding and self-associative precipitation. Because protein solubility can be regarded as a basis for the potential ability to sequester from solvent, we assume that poorly soluble proteins tend not only to precipitate, but also to form solution structures.

12p viettel02 22-02-2013 46 4   Download