A simple fluorescent probe based on a pyrene derivative for rapid detection of protamine and monitoring of trypsin activity.

We report the synthesis of a simple pyrene derivative and its application in protamine detection and monitoring of trypsin activity. This assay can be conducted in aqueous solution and features rapid response, visual detection, high sensitivity and selectivity. The limit of detection of protamine was 0.5 μg mL(-1). The IC50 value of a soybean trypsin inhibitor was estimated to be 0.51 U mL(-1).

[1]  Iván Castelló Serrano,et al.  Dual core quantum dots for highly quantitative ratiometric detection of trypsin activity in cystic fibrosis patients. , 2014, Nanoscale.

[2]  Kyung-Chul Choi,et al.  Risk of cardiovascular disease is suppressed by dietary supplementation with protamine and chitooligosaccharide in Sprague-Dawley rats. , 2013, Molecular medicine reports.

[3]  Masanori Kato,et al.  Plasma α2‐macroglobulin‐trypsin complexlike substance (MTLS) in pancreatic disease , 1996 .

[4]  W. Qin,et al.  Potentiometric determination of trypsin using a polymeric membrane polycation-sensitive electrode based on current-controlled reagent delivery. , 2012, Bioelectrochemistry.

[5]  M. Stevens,et al.  Label-free multimodal protease detection based on protein/perylene dye coassembly and enzyme-triggered disassembly. , 2014, Analytical chemistry.

[6]  Kevin J. Robbins,et al.  K114 (trans, trans)‐bromo‐2,5‐bis(4‐hydroxystyryl)benzene is an efficient detector of cationic amyloid fibrils , 2015, Protein science : a publication of the Protein Society.

[7]  C. Haglund,et al.  Tumour-Associated Trypsin Inhibitor TATI Is a Prognostic Marker in Colorectal Cancer , 2012, Oncology.

[8]  B. Liu,et al.  Anionic conjugated polymer with aptamer-functionalized silica nanoparticle for label-free naked-eye detection of lysozyme in protein mixtures. , 2010, Langmuir : the ACS journal of surfaces and colloids.

[9]  Wenying Li,et al.  Real-time fluorometric assay for acetylcholinesterase activity and inhibitor screening through the pyrene probe monomer-excimer transition. , 2013, Organic letters.

[10]  Shouzhuo Yao,et al.  Gold nanoparticle coupled with fluorophore for ultrasensitive detection of protamine and heparin. , 2013, Talanta.

[11]  Y. Ogata,et al.  Protamine stimulates bone sialoprotein gene expression. , 2013, Gene.

[12]  F. Boulton,et al.  NEONATAL COAGULATION , 1975, The Lancet.

[13]  D. Williams,et al.  Development and validation of an enzyme-linked immunosorbent assay for feline trypsin-like immunoreactivity. , 2000, American journal of veterinary research.

[14]  J. Chambron,et al.  Supramolecular chemical sensors based on pyrene monomer-excimer dual luminescence. , 2011, Chemistry, an Asian journal.

[15]  Shu Pang,et al.  A fluorescence assay for the trace detection of protamine and heparin , 2014 .

[16]  G. Bayramoglu,et al.  Development of a sensitive method for selection of affinity ligand for trypsin using quartz crystal microbalance sensor , 2012, Bioprocess and Biosystems Engineering.

[17]  J. Folkman,et al.  Protamine is an inhibitor of angiogenesis , 1982, Nature.

[18]  Ping Zhang,et al.  Label-free and real-time monitoring of trypsin activity in living cells by quantum-dot-based fluorescent sensors , 2014 .

[19]  M. Heller,et al.  Whole blood assay for trypsin activity using polyanionic focusing gel electrophoresis , 2010, Electrophoresis.

[20]  Yongxin Li,et al.  A fluorescence turn-on method for real-time monitoring of protease activity based on the electron transfer between a fluorophore labeled oligonucleotide and cytochrome c. , 2013, Analytica chimica acta.

[21]  S. Parveen,et al.  Highly sensitive fluorescent detection of trypsin based on BSA-stabilized gold nanoclusters. , 2012, Biosensors & bioelectronics.

[22]  Yuqing Wu,et al.  A unique protein labeling system based on melittin and the non-covalent binding-induced pyrene excimer. , 2010, Chemical communications.

[23]  Wenying Li,et al.  A label-free real time fluorometric assay for protease and inhibitor screening with a released heme. , 2012, Chemical communications.

[24]  Y. Hori,et al.  Development of a fluorogenic probe with a transesterification switch for detection of histone deacetylase activity. , 2012, Journal of the American Chemical Society.

[25]  Ashley C. Gucinski,et al.  Identification of site-specific heterogeneity in peptide drugs using intact mass spectrometry with electron transfer dissociation. , 2014, Rapid communications in mass spectrometry : RCM.

[26]  Yuqing Wu,et al.  A Continuous Fluorometric Assay for Trypsin Based on Melittin and the Noncovalent-binding-induced Pyrene Excimer , 2013 .

[27]  Deqing Zhang,et al.  A new label-free continuous fluorometric assay for trypsin and inhibitor screening with tetraphenylethene compounds. , 2010, Organic letters.

[28]  A. Hvass,et al.  Determination of protamine peptides in insulin drug products using reversed phase high performance liquid chromatography. , 2005, Journal of pharmaceutical and biomedical analysis.

[29]  Ben Zhong Tang,et al.  A dual-mode fluorescence "turn-on" biosensor based on an aggregation-induced emission luminogen. , 2014, Journal of materials chemistry. B.

[30]  Hai‐Chen Wu,et al.  Colorimetric and fluorescent detection of protamines with an anionic polythiophene derivative. , 2013, Organic and biomolecular chemistry.

[31]  Bin Liu,et al.  Conjugated polyelectrolyte based fluorescence turn-on assay for real-time monitoring of protease activity. , 2010, Analytical chemistry.

[32]  K. Suzuki,et al.  Studies on protamines. XVI. The complete amino acid sequence of clupeine YII. , 1972, Journal of biochemistry.

[33]  N. Nishi,et al.  Primary structure of scombrine gamma, protamine isolated from spotted mackerel (Scomber australasicus). , 1993, Journal of biochemistry.

[34]  H. Rinderknecht,et al.  Activation of pancreatic zymogens , 1986, Digestive Diseases and Sciences.

[35]  Lingxin Chen,et al.  Ultrasensitive surface-enhanced Raman scattering detection of trypsin based on anti-aggregation of 4-mercaptopyridine-functionalized silver nanoparticles: an optical sensing platform toward proteases. , 2013, Nanoscale.

[36]  P. Song,et al.  Fluorescence turn-on detection of protamine based on aggregation-induced emission enhancement characteristics of 4-(6'-carboxyl)hexyloxysalicylaldehyde azine. , 2010, The Analyst.

[37]  He Tian,et al.  Insight into aggregation-induced emission characteristics of red-emissive quinoline-malononitrile by cell tracking and real-time trypsin detection , 2014 .

[38]  Masahiko Hirota,et al.  The role of trypsin, trypsin inhibitor, and trypsin receptor in the onset and aggravation of pancreatitis , 2006, Journal of Gastroenterology.

[39]  T. Joo,et al.  γ-Oxo-1-pyrenebutyric acid used for fluorescent detection of serum albumins and trypsin , 2012 .

[40]  Ho-il Choi,et al.  Development of peptide substrates for trypsin based on monomer/excimer fluorescence of pyrene. , 2002, Analytical biochemistry.

[41]  Wenying Li,et al.  Real-time fluorometric turn-on assay for protease activity and inhibitor screening with a benzoperylene probe. , 2014, The Analyst.

[42]  Xiao-yan Li,et al.  A sensitive assay for trypsin using poly(thymine)-templated copper nanoparticles as fluorescent probes. , 2015, The Analyst.

[43]  L. E. Hughes.  TREATMENT OF MALIGNANT DISEASE WITH PROTAMINE SULPHATE. , 1964, Lancet.

[44]  Eric Bakker,et al.  Reversible electrochemical detection of nonelectroactive polyions. , 2003, Journal of the American Chemical Society.