Food and Fermentation Industries >

2019 , Vol. 45 >Issue 19: 17 - 24

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.020989

Research Report

Construction of an in vivo system of Escherichia coli for screening Aβ42 aggregation inhibitors based on β-lactamase

  • ZHAO Wenping ,
  • JIA Longgang ,
  • LU Fuping ,
  • LIU Fufeng
Expand
  • Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, National Engineering Laboratory for Industrial Enzymes,Tianjin Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China

Revised date: 2019-07-12

  Online published: 2019-11-15

Fold

Abstract

Misfolding and aggregation of amyloid β protein (Aβ) play a key role in developing Alzheimer's disease (AD). Therefore, establishing aggregation inhibitors of Aβ is important for alleviating and treating AD. Based on the structural property of TEM1-β-lactamase (βlac) and the aggregation characteristics of Aβ42, this study inserted Aβ42 between residue 196 and 197 of βlac to construct BL21-βlac-Aβ42 recombinant strain. Moreover, it was identified using several known Aβ42 inhibitors. The results showed that with the addition of known Aβ42 inhibitors, the growth of recombinant BL21-βlac-Aβ42 was improved in ampicillin with specified concentration, and the maximum cell dilution for cell growth was also increased. Overall, this system realized high-throughput screening of amyloid aggregation inhibitors in microorganisms, which is helpful for developing new anti-Aβ aggregation drugs.

Key words： Alzheimer’s disease (AD);; amyloid β protein 42 (Aβ42);; TEM1-β-lactamase (βlac);; expression of fusion protein; aggregation inhibitors; in vivo screening system of Escherichia coli

Cite this article

ZHAO Wenping , JIA Longgang , LU Fuping , LIU Fufeng . Construction of an in vivo system of Escherichia coli for screening Aβ42 aggregation inhibitors based on β-lactamase[J]. Food and Fermentation Industries, 2019 , 45(19) : 17 -24 . DOI: 10.13995/j.cnki.11-1802/ts.020989

References

[1] CHITI F, DOBSON C M.Protein misfolding, functional amyloid, and human disease[J]. Annual Review of Biochemistry, 2006, 75(1): 333-366.
[2] CANTER R G, PENNEY J, TSAI L H.The road to restoring neural circuits for the treatment of Alzheimer's disease[J]. Nature, 2016, 539(7 628): 187-196.
[3] HARDY J A, HIGGINS G A.Alzheimer's disease: the amyloid cascade hypothesis[J]. Science, 1992, 256(5 054): 184-185.
[4] 李丽, 刘夫锋. 淀粉样β蛋白质构象转换及其抑制的分子动力学模拟[J]. 生物加工过程, 2019, 17(1): 50-58.
[5] GEERT V G, WIM A.Amyloid, presenilins, and Alzheimer's disease[J]. Neuroscientist A Review Journal Bringing Neurobiology Neurology & Psychiatry, 2003, 9(2): 117-126.
[6] GUO J, SUN W, LIU F F.Brazilin inhibits the Zn2+-mediated aggregation of amyloid β-protein and alleviates cytotoxicity[J]. Journal of Inorganic Biochemistry, 2017, 177: 183-189.
[7] JIA L G, WANG W J, SHANG J Z, et al.Highly efficient soluble expression, purification and characterization of recombinant Ab42 from Escherichia coli[J]. RSC Advances, 2018, 8(33): 18 434-18 441.
[8] LIU F F, WANG W J, SANG J C, et al.Hydroxylated single-walled carbon nanotubes inhibit a beta(42) fibrillogenesis, disaggregate mature fibrils, and protect against a beta(42)-induced cytotoxicity[J]. ACS Chemical Neuroscience, 2019, 10(1): 588-598.
[9] 郑嵋戈, 阮志刚,刘靖,等. β-淀粉样蛋白的代谢机制[J]. 神经解剖学杂志, 2011, 27(3): 344-348.
[10] HARDY J, SELKOE D J.The amyloid hypothesis of Alzheimer's disease: Progress and problems on the road to therapeutics[J]. Science, 2002, 297(5 580): 353-356.
[11] TAPIOLA T, ALAFUZOFF I, HERUKKA S K, et al.Cerebrospinal fluid β-Amyloid 42 and tau proteins as biomarkers of Alzheimer-Type pathologic changes in the brain[J]. Alzheimers & Dementia the Journal of the Alzheimers Association, 2009, 6(4): S170-S170.
[12] COHEN S I, LINSE S, LUHESHI L M, et al.Proliferation of amyloid-β42 aggregates occurs through a secondary nucleation mechanism[J]. Proc Natl Acad Sci USA, 2013, 110(24): 9 758-9 763.
[13] SHANKAR G M, BLOODGOOD B L, TOWNSEND M, et al.Natural oligomers of the alzheimer amyloid-β protein induce reversible synapse loss by modulating an NMDA-Type glutamate receptor-dependent signaling pathway[J]. Journal of Neuroscience, 2007, 27(11): 2 866-2 875.
[14] BIANCALANA M, KOIDE S. Molecular mechanism of Thioflavin-T binding to amyloid fibrils [J]. Biochimica et Biophysica Acta-Proteins and Proteomics, 2010, 1 804(7): 1 405-1 412.
[15] GROENNING M.Binding mode of Thioflavin T and other molecular probes in the context of amyloid fibrils-current status[J]. Journal of Chemical Biology, 2010, 3(1): 1-18.
[16] ZHAO D S, CHEN Y X, LIU Q, et al.Exploring the binding mechanism of thioflavin-T to the β-amyloid peptide by blind docking method[J]. Science China:Chemistry, 2012, 55(1): 112-117.
[17] LI S, LIU F F, YU L L, et al.Dual effect of thioflavin T on the nucleation kinetics of amyloid beta-protein 40[J]. Acta Physico-Chimica Sinica, 2016, 32(6): 1 391-1 396.
[18] BIANCALANA M, MAKABE K, KOIDE A, et al.Molecular mechanism of thioflavin-T binding to the surface of beta-rich peptide self-assemblies[J]. Journal of Molecular Biology, 2009, 385(4): 1 052-1 063.
[19] MENG F, MAREK P, POTTER K J, et al.Rifampicin does not prevent amyloid fibril formation by human islet amyloid polypeptide but does inhibit fibril thioflavin-T interactions: Implications for mechanistic studies of β-cell death[J]. Biochemistry, 2008, 47(22): 6 016-6 024.
[20] OSTERMEIER M.Designing switchable enzymes[J]. Current Opinion in Structural Biology, 2009, 19(4): 442-448.
[21] SAUNDERS J C, YOUNG, L M, MAHOOD R A, et al.An in vivo platform for identifying inhibitors of protein aggregation[J]. Nature Chemical Biology, 2016, 12(2): 94-101.
[22] DELAIRE M, LENFANT F, LABIA R, et al.Site-directed mutagenesis on TEM-1 beta-lactamase: role of Glu166 in catalysis and substrate binding[J]. Protein Engineering, 1991, 4(7): 805-810.
[23] FOIT L, BARDWELL J C.A tripartite fusion system for the selection of protein variants with increased stability in vivo[J]. Methods in Molecular Biology, 2013, 978: 1-20.
[24] ZEBALA J, BARANY F.Mapping catalytically important regions of an enzyme using two-codon insertion mutagenesis: a case study correlating β-lactamase mutants with the three-dimensional structure[J]. Gene, 1991, 100: 51-57.
[25] FOIT L, MORGAN G J, KERN M J, et al.Optimizing protein stability in vivo[J]. Molecular Cell, 2009, 36(5): 861-871.
[26] HALLET B, SHERRATT D J, HAYES F.Pentapeptide scanning mutagenesis: random insertion of a variable five amino acid cassette in a target protein[J]. Nucleic Acids Research, 1997, 25(9): 1 866-1 867.
[27] GALARNEAU A, PRIMEAU M, TRUDEAU L E, et al.Beta-Lactamase protein fragment complementation assays as in vivo and in vitro sensors of protein-protein interactions[J]. Nature Biotechnology, 2002, 20(6): 619-622.
[28] EDWARDS W R, BUSSE K, ALLEMANN R K, et al.Linking the functions of unrelated proteins using a novel directed evolution domain insertion method[J]. Nucleic Acids Research, 2008, 36(13): e78-e78.
[29] GUNTAS G, MANSELL T J, KIM J R, et al.Directed evolution of protein switches and their application to the creation of ligand-binding proteins[J]. Proceedings of the National Academy of Sciences, 2005, 102(32): 11 224-11 229.
[30] YANG F, LIM G P, BEGUM A N, et al.Curcumin inhibits formation of amyloid beta oligomers and fibrils, binds plaques, and reduces amyloid in vivo[J]. Journal of Biological Chemistry, 2005, 280(7): 5 892-5 901.
[31] 刘彬, 赵莹,于思礼, 等. 微生物代谢工程发酵姜黄素研究进展[J]. 食品与发酵工业, 2016, 42(7): 254-260.
Options
Outlines

/

〈 〉

Powered by Beijing Magtech Co. Ltd,.