نتایج جستجو برای: stimulated by his environment

تعداد نتایج: 7350401  

Journal: :The Biochemical journal 1999
C H Klaassen P H Bovee-Geurts G L Decaluwé W J DeGrip

Here we describe a generic procedure for the expression and purification of milligram quantities of functional recombinant eukaryotic integral membrane proteins, exemplified by hexahistidine-tagged bovine rhodopsin. These quantities were obtained with the recombinant baculovirus/Sf9 insect cell-based expression system in large-scale bioreactor cultures with the use of a serum-free and protein-f...

Journal: :Journal of molecular biology 2002
Markus Kaufmann Peter Lindner Annemarie Honegger Kerstin Blank Markus Tschopp Guido Capitani Andreas Plückthun Markus G Grütter

The crystal structure of a mutant form of the single-chain fragment (scFv), derived from the monoclonal anti-His tag antibody 3D5, in complex with a hexahistidine peptide has been determined at 2.7 A resolution. The peptide binds to a deep pocket formed at the interface of the variable domains of the light and the heavy chain, mainly through hydrophobic interaction to aromatic residues and hydr...

Journal: :The Biochemical journal 1993
H Klima A Klein G van Echten G Schwarzmann K Suzuki K Sandhoff

The cDNA of the human GM2-activator protein was cloned into the expression vector pHX17. The plasmid encodes a fusion protein with a hexahistidine tail and a Factor Xa cleavage site at its N-terminus. The recombinant protein was purified from cell homogenates under denaturing conditions by metal-ion affinity chromatography in a single step and then was refolded. The hexahistidine tail could be ...

2017
Dorothee Wasserberg Jordi Cabanas-Danés Jord Prangsma Shane O'Mahony Pierre-Andre Cazade Eldrich Tromp Christian Blum Damien Thompson Jurriaan Huskens Vinod Subramaniam Pascal Jonkheijm

We report oriented immobilization of proteins using the standard hexahistidine (His6)-Ni2+:NTA (nitrilotriacetic acid) methodology, which we systematically tuned to give control of surface coverage. Fluorescence microscopy and surface plasmon resonance measurements of self-assembled monolayers (SAMs) of red fluorescent proteins (TagRFP) showed that binding strength increased by 1 order of magni...

Journal: :Biochemistry 2006
David E Blair Omid Hekmat Alexander W Schüttelkopf Binesh Shrestha Ken Tokuyasu Stephen G Withers Daan M F van Aalten

The fungal pathogen Colletotrichum lindemuthianum secretes an endo-chitin de-N-acetylase (ClCDA) to modify exposed hyphal chitin during penetration and infection of plants. Although a significant amount of biochemical data is available on fungal chitin de-N-acetylases, no structural data exist. Here we describe the 1.8 A crystal structure of a ClCDA product complex and the analysis of the react...

Journal: :Chembiochem : a European journal of chemical biology 2009
Thomas André Annett Reichel Karl-Heinz Wiesmüller Robert Tampé Jacob Piehler Roland Brock

The development of synthetic, low-molecular-weight ligand receptor systems for the selective control of biomolecular interactions remains a major challenge. Binding of oligohistidine peptides to chelators containing Ni2+-loaded nitrilotriacetic acid (NTA) moieties is one of the most widely used and best-characterised recognition systems. Recognition units containing multiple NTA moieties (multi...

Journal: :Methods in molecular biology 2009
Brian P Austin Sreedevi Nallamsetty David S Waugh

Insolubility of recombinant proteins in Escherichia coli is a major impediment to their production for structural and functional studies. One way to circumvent this problem is to fuse an aggregation-prone protein to a highly soluble partner. E. coli maltose-binding protein (MBP) has emerged as one of the most effective solubilizing agents. In this chapter, we describe how to construct combinato...

Journal: :Journal of bacteriology 2005
Julio Ramos Aires Hiroshi Nikaido

To understand better the mechanisms of resistance-nodulation-division (RND)-type multidrug efflux pumps, we examined the Escherichia coli AcrD pump, whose typical substrates, aminoglycosides, are not expected to diffuse spontaneously across the lipid bilayer. The hexahistidine-tagged AcrD protein was purified and reconstituted into unilamellar proteoliposomes. Its activity was measured by the p...

Journal: :Acta crystallographica. Section D, Biological crystallography 2002
David M Duda Craig Yoshioka Lakshmanan Govindasamy Haiqian An Chingkuang Tu David N Silverman Robert McKenna

Carbonic anhydrases catalyze the interconversion of carbon dioxide to bicarbonate. Human carbonic anhydrase isozyme III with a C-terminal hexahistidine tag was overexpressed in Eschericha coli, purified and crystallized. Diffraction data (93.4% completeness) were collected to 2.2 A resolution on an in-house R-AXIS IV++ image-plate system with Osmic mirrors and a Rigaku HU-H3R CU rotating-anode ...

2016
Alexander Kinna Berend Tolner Enrique Miranda Rota Nigel Titchener‐Hooker Darren Nesbeth Kerry Chester

Fusion-tag affinity chromatography is a key technique in recombinant protein purification. Current methods for protein recovery from mammalian cells are hampered by the need for feed stream clarification. We have developed a method for direct capture using immobilized metal affinity chromatography (IMAC) of hexahistidine (His6) tagged proteins from unclarified mammalian cell feed streams. The p...

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