title: Study on nanocomposite construction based on the multi-functional biotemplate self-assembled by the recombinant TMGMV coat protein for potential biomedical applications.
abstract: Nowadays there is a growing interest in bio-scaffolded nanoarchitectures. Rapid progress in nanobiotechnology and molecular biology has allowed the engineering of inorganic-binding peptides termed as genetically engineered polypeptides for inorganics (GEPIs) into self-assembling biological structures to facilitate the design of novel biomedical or bioimaging devices. Here we introduce a novel nanocomposite comprising a self-assembled protein scaffold based on a recombinant tobacco mild green mosaic tobamovirus (TMGMV) coat protein (CP) and the photocatalytic TiO2 nanoparticles attached to it, which may provide a generic method for materials engineering. A template containing a modified TMGMV CP (mCP) gene, with the first six C-terminal amino acid residues deleted to accommodate more foreign peptides and expressing a site-directed mutation of A123C for bioconjugation utility, and two genetically engineered mutants, Escherichia coli-based P-mCP-Ti7 containing a C-terminal TiO2 GEPI sequence of seven peptides (Ti7) and Hi5 insect cells-derived E-CP-Ti7-His6 C-terminally fused with Ti7+His6 tag were created. Expression vectors and protocols for enriching of the two CP variants were established and the resultant proteins were identified by western blot analysis. Their RNA-free self-assembling structures were analyzed by transmission electron microscopy (TEM) and immuno-gold labeling TEM analysis. Adherence of nanoparticles to the P-mCP-Ti7 induced protein scaffold was visualized by TEM analysis. Also discussed is the Cysteine thiol reactivity in bioconjugation reactions with the maleimide-functionalized porphyrin photosensitizers which can function as clinical photodynamic therapy agents. This study introduced a novel approach to producing an assembly-competent recombinant TMGMV CP, examined its ability to serve as a novel platform for the multivalent display of surface ligands and demonstrated an alternative method for nanodevice synthesis for nanobiotechnological applications by combining GEPIs-mediated immobilization with the controllability of self-assembling recombinant TMGMV CP.
authors: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, People's Republic of China.
authors: Lei
authors: L
authors: Song
authors: Shiwen
authors: S
authors: Wang
authors: Haina
authors: H
authors: Wang
authors: Hua
authors: H
authors: Zhang
authors: Haolong
authors: H
authors: Cong
authors: Xingyu
authors: X
authors: Jiang
authors: Po
authors: P
authors: Tien
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000737:chemistry* / Q000235:genetics; D002460:Cell Line; D053768:Metal Nanoparticles / Q000737:chemistry / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission; D053761:Nanocomposites / Q000737:chemistry* / Q000648:ultrastructure; D036103:Nanotechnology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D014025:Titanium / Q000737:chemistry; D014027:Tobacco Mosaic Virus / Q000737:chemistry* / Q000235:genetics
chemical_list: D036022:Capsid Proteins; C009496:H protein, Tobacco mosaic virus; D011994:Recombinant Proteins; C009495:titanium dioxide; D014025:Titanium
references: Annu Rev Phytopathol. 2008;46:361-84
references: 18473700
references: J Virol Methods. 2013 May;189(2):328-40
references: 23499261
references: Curr Opin Biotechnol. 2004 Dec;15(6):513-7
references: 15560977
references: Nat Biotechnol. 1997 Mar;15(3):269-72
references: 9062928
references: Biosens Bioelectron. 2007 Apr 15;22(9-10):1841-52
references: 17071070
references: Biophys J. 2006 Aug 15;91(4):1501-12
references: 16731551
references: Nano Lett. 2005 Jul;5(7):1429-34
references: 16178252
references: J Mol Biol. 1992 Nov 20;228(2):516-28
references: 1453461
references: Toxicol Lett. 2009 Sep 28;189(3):191-9
references: 19486931
references: Science. 2002 May 3;296(5569):892-5
references: 11988570
references: Nat Mater. 2010 Aug;9(8):676-84
references: 20651808
references: J Am Chem Soc. 2003 Apr 23;125(16):4700-1
references: 12696875
references: Acta Biomater. 2007 May;3(3):289-99
references: 17257913
references: ACS Nano. 2011 Mar 22;5(3):1606-16
references: 21361370
references: J Am Chem Soc. 2007 Mar 21;129(11):3104-9
references: 17319656
references: Angew Chem Int Ed Engl. 2006 Nov 27;45(46):7754-8
references: 17072921
references: Nat Mater. 2010 Feb;9(2):165-71
references: 19946279
references: Science. 1998 Sep 25;281(5385):2013-6
references: 9748157
references: Bioelectrochemistry. 2007 Nov;71(2):217-22
references: 17643355
references: Angew Chem Int Ed Engl. 2007;46(18):3184-93
references: 17348058
references: Nat Mater. 2005 Jun;4(6):435-46
references: 15928695
references: J Nanosci Nanotechnol. 2002 Feb;2(1):95-100
references: 12908327
references: Nano Lett. 2010 Mar 10;10(3):773-6
references: 20158260
references: Adv Drug Deliv Rev. 2012 Jun 15;64(9):811-25
references: 22285585
references: Biotechnol Bioeng. 2012 Jan;109(1):16-30
references: 21915854
references: J Nanosci Nanotechnol. 2010 Dec;10(12):7897-905
references: 21121277
references: Bioconjug Chem. 2008 Sep;19(9):1785-95
references: 18681468
references: Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):9067-71
references: 8090770
references: Small. 2009 Aug 3;5(15):1706-21
references: 19572330
references: Nanoscale. 2011 Mar;3(3):1004-7
references: 21132192
references: J Virol Methods. 2010 Jun;166(1-2):77-85
references: 20219539
references: Nat Mater. 2003 Sep;2(9):577-85
references: 12951599
references: Protein Expr Purif. 2006 Jun;47(2):490-7
references: 16406677
references: Nano Lett. 2007 Aug;7(8):2407-16
references: 17630812
references: Biotechnology (N Y). 1995 Oct;13(10):1079-84
references: 9636281
references: Anal Chem. 2002 Feb 15;74(4):841-7
references: 11866065
references: Angew Chem Int Ed Engl. 2003;42(47):5796-800
references: 14673909
references: Langmuir. 2004 Aug 31;20(18):7720-8
references: 15323524
references: Science. 2004 Jan 9;303(5655):213-7
references: 14716009
references: Protein Sci. 1999 Nov;8(11):2418-23
references: 10595544
references: J Am Chem Soc. 2011 Jul 13;133(27):10482-9
references: 21627173
references: Nature. 1996 Aug 15;382(6592):607-9
references: 8757129
references: Biophys J. 1980 Oct;32(1):460-2
references: 19431395
references: Phys Chem Chem Phys. 2013 Apr 14;15(14):4844-58
references: 23450160
references: Proc Natl Acad Sci U S A. 2011 Oct 11;108(41):16998-7003
references: 21949376
references: Mutat Res. 2007 Apr 2;628(2):99-106
references: 17223607
references: Vaccine. 2006 Jan 12;24(2):109-15
references: 16337317
references: Curr Top Microbiol Immunol. 2009;327:95-122
references: 19198572