Search results for "biotin"

Purification by affinity chromatography of H1 RNA-Binding Proteins from rat brain

2003

Post-transcriptional regulation of mRNA metabolism is involved in processes as different as cell fate specification in development and cell response to a large variety of environmental cues. Regulation of all steps of RNA metabolism depends on RNA-binding proteins (RBPs). By using a T1 RNase protection assay, we previously identified three H1° RNA-binding factors (p40, p70 and p110), highly expressed in the rat brain. Here we report enrichment of these factors from brain extracts, obtained by affinity chromatography of biotinylated H1° RNA-protein complexes on streptavidin-conjugated paramagnetic particles. The purified proteins maintain RNA-binding ability and preference for histone messag…

Efficient gene therapy based targeting system for the treatment of inoperable tumors

2012

Background A considerable percentage of tumors are not amenable to surgery. We have designed a simple and powerful targeting system that offers an alternative option for the multi-component pre-targeting strategies used clinically. This targeting system can be used for any type of solid tumors independent of the tumor type, thereby omitting the need to engineer unique antibodies for each specific application or tumour type. In the present study, we show the expression of a chimeric fusion protein, which contains the low-density lipoprotein receptor transmembrane domains and avidin, after local gene transfer and its ability to bind biotinylated compounds in vivo. Methods Semliki Forest virus…

Chicken genome analysis reveals novel genes encoding biotin-binding proteins related to avidin family

2005

Background A chicken egg contains several biotin-binding proteins (BBPs), whose complete DNA and amino acid sequences are not known. In order to identify and characterise these genes and proteins we studied chicken cDNAs and genes available in the NCBI database and chicken genome database using the reported N-terminal amino acid sequences of chicken egg-yolk BBPs as search strings. Results Two separate hits showing significant homology for these N-terminal sequences were discovered. For one of these hits, the chromosomal location in the immediate proximity of the avidin gene family was found. Both of these hits encode proteins having high sequence similarity with avidin suggesting that chic…

Structure and characterization of a novel chicken biotin-binding protein A (BBP-A)

2007

Background. The chicken genome contains a BBP-A gene showing similar characteristics to avidin family genes. In a previous study we reported that the BBP-A gene may encode a biotin-binding protein due to the high sequence similarity with chicken avidin, especially at regions encoding residues known to be located at the ligand-binding site of avidin. Results. Here, we expand the repertoire of known macromolecular biotin binders by reporting a novel biotin-binding protein A (BBP-A) from chicken. The BBP-A recombinant protein was expressed using two different expression systems and purified with affinity chromatography, biochemically characterized and two X-ray structures were solved – in comp…

Functionalized carbon nanotube for next generation biosensor

2013

Thioredoxin-related protein of 14 kDa may directly reduce protein cysteinylation motifs

2018

Disulfide stress has been associated with inflammation and characterized by an increase in cystine levels and protein cysteinylation. Furthermore, it was recently discovered that thioredoxin-related protein of 14 kDa (TRP14, encoded by TXNDC17) exhibits efficient cystine reductase activity. The aim of our research was to elucidate if TRP14 is also able to reduce cysteinylated proteins in mammalian cells. Thus, protein cysteinylation was assessed in control and TRP14 knockdown cells in vitro through their pre-treatment with 25 µg/ml cycloheximide for 30 min and incubation with 250 µM biotinylated cysteine for 1 h. Moreover, such TRP14 knockdown cell lysates were tested as cysteinylated subst…

Reversed-phase liquid chromatography of biotin-labelled nucleotides

1985

Determination of the stability of protein pools from the cell wall of fungi.

2002

Stability of the protein populations present in the cell wall of three ascomycetous fungi Candida albicans, Saccharomyces cerevisiae and Yarrowia lipolytica was investigated. Cell wall proteins were either labeled with biotin or radiolabeled with amino acids, and chased for a period of time representing several generations. Proteins linked by non-covalent or covalent bonds were separated and their turnover was analyzed. No significant turnover took place during the chase period, and in fact radioactive proteins were accumulated in the wall during the period possibly by transfer through the secretory pathway. This transfer did not involve de novo protein synthesis; it was inhibited by azide,…

Occurrence of three different binding sites forBacillus thuringiensisδ-endotoxins in the midgut brush border membrane of the potato tuber moth,phthor…

1994

The potato tuber moth is susceptible to at least three insecticidal crystal proteins (ICPs) from Bacillus thuringiensis: CrylA(b), CrylB, and CrylC. To design useful combinations of toxin genes either in transgenic plants or in new genetically modified B. thuringiensis strains, it is necessary to determine the binding characteristics of the different ICPs so as not to combine a pair sharing the same binding site. This has been accomplished using two different techniques: 125I-labeling of the ICPs with further measurement of the radioactivity bound to brush border membrane vesicles, and microscopic visualization of the bound ICPs by enzyme-linked reagents such as antibodies or streptavidin u…

Nanodevices by DNA based gold nanostructures

2017

In this thesis DNA based structures were utilized to create gold nanostructures for nanosensing and nanoelectronic applications. In the past, both of these fields have been dominated by the conventional lithography methods, e.g., electron beam lithography and UV-lithography, but more recently scaling down the components by these techniques has become increasingly more complex and costly. Especially in the micro- and nanoelectronics, the increase in the component density and thus computational power would require fabrication of sub-10-nm components, which is challenging for the top-down approaches. Aforementioned developments have led researchers to seek alternative methods to fabricate these…