Oriented immobilisation of histidine-tagged protein and its application in exploring interactions between ligands and proteins View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2014-05

AUTHORS

Xinfeng Zhao, Qian Li, Chaoni Xiao, Yajun Zhang, Liujiao Bian, Jianbin Zheng, Xiaohui Zheng, Zijian Li, Youyi Zhang, Taiping Fan

ABSTRACT

A method based on reaction with a diazonium salt was developed to immobilise oriented His-tagged protein onto silica gel. The binding efficiency of the phenylamine-group-coated gel was determined to be 65 %, providing a binding capacity of His-tagged protein up to the gram level. Using His-tagged β2-adrenoceptor (β2-AR) as a probe, we developed a new mathematical model to elucidate the interactions between the receptor and five ligands (methoxyphenamine, terbutaline, salbutamol, tulobuterol and fenoterol). These drugs proved to only have one type of binding site on the immobilised β2-AR, yielding higher association constants and numbers of binding sites than random attachment assays. The association constants determined by the new model positively correlated to the values from a radioligand binding method, with a regression equation of y = 1.75x - 7.18 and a correlation coefficient of 0.9807. The oriented method resulted in a high binding capacity and quantitative immobilisation of the His-tagged protein. The proposed model can be used to determine the interactions between the ligands and the immobilised protein with the advantages of drug and time saving. More... »

PAGES

2975-2985

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s00216-014-7723-x

DOI

http://dx.doi.org/10.1007/s00216-014-7723-x

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1019103072

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/24633512


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Download the RDF metadata as:  json-ld nt turtle xml License info

HOW TO GET THIS DATA PROGRAMMATICALLY:

JSON-LD is a popular format for linked data which is fully compatible with JSON.

curl -H 'Accept: application/ld+json' 'https://scigraph.springernature.com/pub.10.1007/s00216-014-7723-x'

N-Triples is a line-based linked data format ideal for batch operations.

curl -H 'Accept: application/n-triples' 'https://scigraph.springernature.com/pub.10.1007/s00216-014-7723-x'

Turtle is a human-readable linked data format.

curl -H 'Accept: text/turtle' 'https://scigraph.springernature.com/pub.10.1007/s00216-014-7723-x'

RDF/XML is a standard XML format for linked data.

curl -H 'Accept: application/rdf+xml' 'https://scigraph.springernature.com/pub.10.1007/s00216-014-7723-x'


 

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