Engineering PQQ-glucose dehydrogenase into an allosteric electrochemical Ca2+ sensor

被引:33
作者
Guo, Zhong [1 ]
Johnston, Wayne A. [1 ]
Stein, Viktor [1 ]
Kalimuthu, Palraj [2 ]
Perez-Alcala, Siro [3 ]
Bernhardt, Paul V. [2 ]
Alexandrov, Kirill [1 ]
机构
[1] Univ Queensland, Inst Mol Biosci, Brisbane, Qld 4072, Australia
[2] Univ Queensland, Sch Chem & Mol Biosci, Brisbane, Qld 4072, Australia
[3] Mol Warehouse Ltd, Twickenham TW1 3DY, England
基金
澳大利亚国家健康与医学研究理事会; 澳大利亚研究理事会;
关键词
PYRROLOQUINOLINE-QUINONE; ACINETOBACTER-CALCOACETICUS; PROTEIN SWITCHES; CALMODULIN; CALCIUM; RECONSTITUTION; INDICATORS; BIOSENSORS; DESIGN;
D O I
10.1039/c5cc07824e
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Electrochemical biosensors convert biological events to an electrical current. To date most electrochemical biosensors exploit activities of naturally occurring enzymes. Here we demonstrated that insertion of a calmodulin domain into the redox enzyme PQQ-glucose dehydrogenase resulted in a selective Ca2+ biosensor that could be used to rapidly measure Ca2+ concentrations in human biological fluids. We were able to convert a point-of-care glucometer into Ca2+ monitor by refurbishing it with the developed biosensor. We propose that similar engineering strategies may be used to create highly specific electrochemical biosensors to other analytes. Compatibility with cheap and ubiquitous amperometric detectors is expected to accelerate progression of these biosensors into clinical applications.
引用
收藏
页码:485 / 488
页数:4
相关论文
共 26 条
  • [1] Screening of Peptide Ligands for Pyrroloquinoline Quinone Glucose Dehydrogenase Using Antagonistic Template-Based Biopanning
    Abe, Koichi
    Yoshida, Wataru
    Terada, Kotaro
    Yagi-Ishii, Yukiko
    Ferri, Stefano
    Ikebukuro, Kazunori
    Sode, Koji
    [J]. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2013, 14 (12): : 23244 - 23256
  • [2] Directed evolution of protein switches and their application to the creation of ligand-binding proteins
    Guntas, G
    Mansell, TJ
    Kim, JR
    Ostermeier, M
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2005, 102 (32) : 11224 - 11229
  • [3] Hu Jamie, 2011, J Diabetes Sci Technol, V5, P1294
  • [4] Construction and characterization of mutant water-soluble PQQ glucose dehydrogenases with altered Km values -: Site-directed mutagenesis studies on the putative active site
    Igarashi, S
    Ohtera, T
    Yoshida, H
    Witarto, AB
    Sode, K
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1999, 264 (03) : 820 - 824
  • [5] Molecular engineering of PQQGDH and its applications
    Igarashi, S
    Okuda, J
    Ikebukuro, K
    Sode, K
    [J]. ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 2004, 428 (01) : 52 - 63
  • [6] Engineered allosteric activation of kinases in living cells
    Karginov, Andrei V.
    Ding, Feng
    Kota, Pradeep
    Dokholyan, Nikolay V.
    Hahn, Klaus M.
    [J]. NATURE BIOTECHNOLOGY, 2010, 28 (07) : 743 - U1756
  • [7] Genetically encoded Ca2+ indicators: Properties and evaluation
    Koldenkova, Vadim Perez
    Nagai, Takeharu
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH, 2013, 1833 (07): : 1787 - 1797
  • [8] The many structural faces of calmodulin: a multitasking molecular jackknife
    Kursula, Petri
    [J]. AMINO ACIDS, 2014, 46 (10) : 2295 - 2304
  • [9] An Engineered Calmodulin-Based Allosteric Switch for Peptide Biosensing
    Meister, Glenna E.
    Joshi, Neel S.
    [J]. CHEMBIOCHEM, 2013, 14 (12) : 1460 - 1467
  • [10] Production, characterization, and reconstitution of recombinant quinoprotein glucose dehydrogenase (soluble type; EC 1.1.99.17) apoenzyme of Acinetobacter calcoaceticus
    Olsthoorn, AJJ
    Duine, JA
    [J]. ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1996, 336 (01) : 42 - 48