Self-assembly-induced far-red/near-infrared fluorescence light-up for detecting and visualizing specific protein-Peptide interactions.

Understanding specific protein-peptide interactions could offer a deep insight into the development of therapeutics for many human diseases. In this work, we designed and synthesized a far-red/near-infrared (FR/NIR) fluorescence light-up probe (DBT-2EEGWRESAI) by simply integrating two tax-interacting protein-1 (TIP-1)-specific peptide ligands (EEGWRESAI) with one 4,7-di(thiophen-2-yl)-2,1,3-benzothiadiazole (DBT) unit. We first demonstrated that DBT is an environment-sensitive fluorophore with FR/NIR fluorescence due to its strong charge transfer character in the excited state. Thanks to the environmental sensitivity of DBT, the probe DBT-2EEGWRESAI is very weakly fluorescent in aqueous solution but lights up its fluorescence when the probe specifically binds to TIP-1 protein or polyprotein (ULD-TIP-1 tetramer). It is found that the DBT-2EEGWRESAI/TIP-1 protein and the DBT-2EEGWRESAI/ULD-TIP-1 tetramer could self-assemble into spherical nanocomplexes and a nanofiber network, respectively, which lead to probe fluorescence turn-on through providing DBT with a hydrophobic microenvironment. By virtue of the self-assembly-induced FR/NIR fluorescence turn-on, DBT-2EEGWRESAI can detect and visualize specific protein/polyprotein-peptide interactions in both solution and live bacteria in a high contrast and selective manner.

[1]  K. Tan,et al.  Environment-sensitive fluorescent turn-on probes targeting hydrophobic ligand-binding domains for selective protein detection. , 2013, Angewandte Chemie.

[2]  P. Jemth,et al.  Probing the role of backbone hydrogen bonds in protein-peptide interactions by amide-to-ester mutations. , 2013, Journal of the American Chemical Society.

[3]  D. Ding,et al.  Imaging: Conjugated Polymer Amplified Far‐Red/Near‐Infrared Fluorescence from Nanoparticles with Aggregation‐Induced Emission Characteristics for Targeted In Vivo Imaging (Adv. Healthcare Mater. 3/2013) , 2013 .

[4]  K. Hahn,et al.  Environment-sensing merocyanine dyes for live cell imaging applications. , 2013, Bioconjugate chemistry.

[5]  Ryu J. Iwatate,et al.  Rational design of highly sensitive fluorescence probes for protease and glycosidase based on precisely controlled spirocyclization. , 2013, Journal of the American Chemical Society.

[6]  Bing Xu,et al.  Imaging enzyme-triggered self-assembly of small molecules inside live cells , 2012, Nature Communications.

[7]  Itaru Hamachi,et al.  Specific cell surface protein imaging by extended self-assembling fluorescent turn-on nanoprobes. , 2012, Journal of the American Chemical Society.

[8]  Cristina Airoldi,et al.  Versatile and efficient targeting using a single nanoparticulate platform: application to cancer and Alzheimer's disease. , 2012, ACS nano.

[9]  Jong Seung Kim,et al.  Direct fluorescence monitoring of the delivery and cellular uptake of a cancer-targeted RGD peptide-appended naphthalimide theragnostic prodrug. , 2012, Journal of the American Chemical Society.

[10]  Ben Zhong Tang,et al.  Specific detection of integrin αvβ3 by light-up bioprobe with aggregation-induced emission characteristics. , 2012, Journal of the American Chemical Society.

[11]  Huaimin Wang,et al.  Rational design of a tetrameric protein to enhance interactions between self-assembled fibers gives molecular hydrogels. , 2012, Angewandte Chemie.

[12]  Juan Li,et al.  General approach for monitoring peptide-protein interactions based on graphene-peptide complex. , 2011, Analytical chemistry.

[13]  Anna Waller,et al.  Simultaneous in vitro molecular screening of protein-peptide interactions by flow cytometry, using six Bcl-2 family proteins as examples , 2011, Nature Protocols.

[14]  D. Chiu,et al.  Near-infrared fluorescent dye-doped semiconducting polymer dots. , 2011, ACS nano.

[15]  M. Gross,et al.  Protein-peptide affinity determination using an H/D exchange dilution strategy: Application to antigen-antibody interactions , 2010, Journal of the American Society for Mass Spectrometry.

[16]  P. Choyke,et al.  New strategies for fluorescent probe design in medical diagnostic imaging. , 2010, Chemical reviews.

[17]  M. Han,et al.  Tumor targeting chitosan nanoparticles for dual-modality optical/MR cancer imaging. , 2010, Bioconjugate chemistry.

[18]  Yan-Kai Tzeng,et al.  Sub‐20‐nm Fluorescent Nanodiamonds as Photostable Biolabels and Fluorescence Resonance Energy Transfer Donors , 2010, Advanced materials.

[19]  B. Imperiali,et al.  Monitoring protein interactions and dynamics with solvatochromic fluorophores. , 2010, Trends in biotechnology.

[20]  Hao Zhou,et al.  Molecular mechanism of inward rectifier potassium channel 2.3 regulation by tax-interacting protein-1. , 2009, Journal of molecular biology.

[21]  B. Imperiali,et al.  A general screening strategy for peptide-based fluorogenic ligands: probes for dynamic studies of PDZ domain-mediated interactions. , 2009, Journal of the American Chemical Society.

[22]  R. Tampé,et al.  Conformation of receptor adopted upon interaction with virus revealed by site-specific fluorescence quenchers and FRET analysis. , 2009, Journal of the American Chemical Society.

[23]  Changfeng Wu,et al.  Multicolor conjugated polymer dots for biological fluorescence imaging. , 2008, ACS nano.

[24]  R. Russell,et al.  Peptide-mediated interactions in biological systems: new discoveries and applications. , 2008, Current opinion in biotechnology.

[25]  Ronald T Raines,et al.  Bright ideas for chemical biology. , 2008, ACS chemical biology.

[26]  D. Lawrence,et al.  Seeing Is Believing: Peptide-Based Fluorescent Sensors of Protein Tyrosine Kinase Activity , 2007 .

[27]  B. Imperiali,et al.  Fluorogenic probes for monitoring peptide binding to class II MHC proteins in living cells. , 2007, Nature chemical biology.

[28]  D. Lawrence,et al.  Seeing Is Believing: Peptide‐Based Fluorescent Sensors of Protein Tyrosine Kinase Activity , 2007, Chembiochem : a European journal of chemical biology.

[29]  Angel W. Lee,et al.  Multiplexed detection of protein-peptide interaction and inhibition using capillary electrophoresis. , 2007, Analytical chemistry.

[30]  Günther Jung,et al.  A fluorescence-based synthetic LPS sensor. , 2007, Journal of the American Chemical Society.

[31]  Timothy Londergan,et al.  Looking towards label-free biomolecular interaction analysis in a high-throughput format: a review of new surface plasmon resonance technologies. , 2006, Current opinion in biotechnology.

[32]  Thierry Livache,et al.  Clinically related protein-peptide interactions monitored in real time on novel peptide chips by surface plasmon resonance imaging. , 2006, Clinical chemistry.

[33]  K. Plaxco,et al.  Engineering a signal transduction mechanism for protein-based biosensors. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[34]  Thomas J Magliery,et al.  Detecting protein-protein interactions with a green fluorescent protein fragment reassembly trap: scope and mechanism. , 2005, Journal of the American Chemical Society.

[35]  K. Hahn,et al.  Activation of Endogenous Cdc42 Visualized in Living Cells , 2004, Science.

[36]  C. Ross,et al.  Protein aggregation and neurodegenerative disease , 2004, Nature Medicine.

[37]  B. Imperiali,et al.  Versatile fluorescence probes of protein kinase activity. , 2003, Journal of the American Chemical Society.

[38]  J. Frangioni In vivo near-infrared fluorescence imaging. , 2003, Current opinion in chemical biology.

[39]  K. Hahn,et al.  Solvent-sensitive dyes to report protein conformational changes in living cells. , 2003, Journal of the American Chemical Society.

[40]  Jürgen Wolfrum,et al.  Detection of individual p53-autoantibodies by using quenched peptide-based molecular probes. , 2002, Angewandte Chemie.

[41]  Junbiao Peng,et al.  Novel red-emitting fluorene-based copolymers , 2002 .

[42]  T R Hughes,et al.  Genetic selection of peptide inhibitors of biological pathways. , 1999, Science.

[43]  S. Fields,et al.  Protein-peptide interactions analyzed with the yeast two-hybrid system. , 1995, Nucleic acids research.