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Heterogeneous sensing of post-translational modification enzymes by integrating the advantage of homogeneous analysis.
Xia, Ning; Sun, Ting; Liu, Lin; Tian, Linxu; Sun, Zhifang.
Afiliação
  • Xia N; Henan Province of Key Laboratory of New Optoelectronic Functional Materials, Anyang Normal University, Anyang, Henan, 455000, PR China.
  • Sun T; Henan Province of Key Laboratory of New Optoelectronic Functional Materials, Anyang Normal University, Anyang, Henan, 455000, PR China; School of Chemistry and Materials Science, Guizhou Education University, GaoXin Road 115, Wudang District, Guizhou, 550000, PR China.
  • Liu L; Henan Province of Key Laboratory of New Optoelectronic Functional Materials, Anyang Normal University, Anyang, Henan, 455000, PR China. Electronic address: liulin@aynu.edu.cn.
  • Tian L; Henan Province of Key Laboratory of New Optoelectronic Functional Materials, Anyang Normal University, Anyang, Henan, 455000, PR China.
  • Sun Z; Henan Province of Key Laboratory of New Optoelectronic Functional Materials, Anyang Normal University, Anyang, Henan, 455000, PR China. Electronic address: allensune@gmail.com.
Talanta ; 237: 122949, 2022 Jan 15.
Article em En | MEDLINE | ID: mdl-34736675
ABSTRACT
Heterogeneous analysis has great application prospects in the detection of post-translational modification (PTM) enzymes with the advantages of signal enhancement, less sample demand, and high sensitivity and selectivity. Nevertheless, once the substrate was fixed on a solid interface, the steric hindrance might limit the approaching of catalytic center to the substrate, thus reducing the efficiency of PTM. Herein, we suggested that the avidin-modified interface could be used to develop heterogeneous sensing platforms with biotin-labeled substrates as the probes, in which the enzymatic PTM was performed in solution and the heterogeneous assay was conducted on a solid surface. The sensing strategy integrates the advantages but overcomes the defects of both homogeneous and heterogeneous assays. Protein kinase A (PKA) and histone acetyltransferase (HAT) were determined as the examples by using sequence-specific peptide substrates. The signal changes were monitored by HRP-based colorimetric assay and antibody-amplified surface plasmon resonance (SPR). The methods were used for analysis of cell lysates and evaluation of inhibition efficiency with satisfactory results. The strategy can be used for the detection of a variety of biological enzymes and provide a new idea for the design of various heterogeneous biosensors. Thus, this work should be of great significance to the popularization and practical application of biosensors.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Técnicas Biossensoriais Idioma: En Revista: Talanta Ano de publicação: 2022 Tipo de documento: Article País de publicação: HOLANDA / HOLLAND / NETHERLANDS / NL / PAISES BAJOS / THE NETHERLANDS

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Técnicas Biossensoriais Idioma: En Revista: Talanta Ano de publicação: 2022 Tipo de documento: Article País de publicação: HOLANDA / HOLLAND / NETHERLANDS / NL / PAISES BAJOS / THE NETHERLANDS