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DOI:	10.1371/journal.pone.0087995
论文题名:	Specific and Reversible Immobilization of Proteins Tagged to the Affinity Polypeptide C-LytA on Functionalized Graphite Electrodes
作者:	Daniel Bello-Gil; Beatriz Maestro; Jennifer Fonseca; Juan M. Feliu; Víctor Climent; Jesús M. Sanz
刊名:	PLOS ONE
ISSN:	1932-6203
出版年:	2014
发表日期:	2014-1-31
卷:	9, 期:1
语种:	英语
英文关键词:	Graphite ; Cholines ; Sodium phosphate ; Electrode potentials ; Diazo compounds ; Electrolytes ; Enzyme immobilization ; Enzymes
英文摘要:	We have developed a general method for the specific and reversible immobilization of proteins fused to the choline-binding module C-LytA on functionalized graphite electrodes. Graphite electrode surfaces were modified by diazonium chemistry to introduce carboxylic groups that were subsequently used to anchor mixed self-assembled monolayers consisting of N,N-diethylethylenediamine groups, acting as choline analogs, and ethanolamine groups as spacers. The ability of the prepared electrodes to specifically bind C-LytA-tagged recombinant proteins was tested with a C-LytA-β-galactosidase fusion protein. The binding, activity and stability of the immobilized protein was evaluated by electrochemically monitoring the formation of an electroactive product in the enzymatic hydrolysis of the synthetic substrate 4-aminophenyl β-D-galactopyranoside. The hybrid protein was immobilized in an specific and reversible way, while retaining the catalytic activity. Moreover, these functionalized electrodes were shown to be highly stable and reusable. The method developed here can be envisaged as a general, immobilization procedure on the protein biosensor field.
URL:	http://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0087995&type=printable
Citation statistics:
资源类型:	期刊论文
标识符:	http://119.78.100.158/handle/2HF3EXSE/18933
Appears in Collections:	过去全球变化的重建影响、适应和脆弱性科学计划与规划气候变化与战略全球变化的国际研究计划气候减缓与适应气候变化事实与影响

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File Name/ File Size	Content Type	Version	Access	License
journal.pone.0087995.PDF（601KB）	期刊论文	作者接受稿	开放获取	CC BY-NC-SA	View Download

作者单位:

Instituto de Biología Molecular y Celular, Universidad Miguel Hernández, Elche, Spain;Instituto Universitario de Electroquímica. Universidad de Alicante, Alicante, Spain;Instituto Universitario de Electroquímica. Universidad de Alicante, Alicante, Spain;Instituto de Biología Molecular y Celular, Universidad Miguel Hernández, Elche, Spain;Instituto Universitario de Electroquímica. Universidad de Alicante, Alicante, Spain;Instituto Universitario de Electroquímica. Universidad de Alicante, Alicante, Spain;Instituto de Biología Molecular y Celular, Universidad Miguel Hernández, Elche, Spain

Recommended Citation:

Daniel Bello-Gil,Beatriz Maestro,Jennifer Fonseca,et al. Specific and Reversible Immobilization of Proteins Tagged to the Affinity Polypeptide C-LytA on Functionalized Graphite Electrodes[J]. PLOS ONE,2014-01-01,9(1)

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