PMID- 27670785
OWN - NLM
STAT- Publisher
LR  - 20191120
IS  - 1756-0500 (Electronic)
IS  - 1756-0500 (Linking)
VI  - 9
IP  - 1
DP  - 2016 Sep 26
TI  - Infrared-emitting, peptidase-resistant fluorescent ligands of the bradykinin B(2) 
      receptor: application to cytofluorometry and imaging.
PG  - 452
LID - 452
AB  - BACKGROUND: We have previously reported the design, pharmacological properties 
      and imaging application of bradykinin (BK) B(2) receptor (B(2)R) ligands 
      conjugated with fluorophores such as fluorescein derivatives at their N-terminus. 
      To take advantage of the high penetration of infrared light into living tissues 
      and their low autofluorescence in this region of the spectrum, additional probes 
      conjugated with cyanine dye 7 (Cy7) were synthesized and characterized. RESULTS: 
      The antagonist B-9430 (D-Arg-[Hyp(3),Igl(5),D-Igl(7),Oic(8)]-BK) and the agonist 
      B-9972 (D-Arg-[Hyp(3),Igl(5),Oic(7),Igl(8)]-BK) were N-terminally extended with 
      the infrared fluorophore Cy7, producing the peptides B-10665 and B-10666, 
      respectively. Pharmacological studies indicated that the agonist B-10666 lost 
      much affinity for the B(2)R vs. the parent peptide, whereas the antagonist 
      B-10665 better retained its potency vs. B-9430 (competition of [(3)H]BK binding 
      to human B(2)R, contractility of the human isolated umbilical vein for which 
      potency losses were more important in each case). Both probes stained HEK 293 
      cells that expressed the B(2)R-green fluorescent protein (GFP) construction in a 
      specific manner (confocal microscopy) and with very extensive co-localization of 
      the green and infrared fluorescence in either case. The agonist B-10666 at 100 nM 
      promoted the endocytosis of B(2)R-GFP in live cells, but not the antagonist 
      version at 10-25 nM. The Cy7-labeled peptides did not label cells expressing the 
      beta(2)-adrenoceptor-GFP construction. B-10665 at low nanomolar concentrations was 
      an effective probe for the recombinant B(2)Rs in cytofluorometry and macroscopic 
      imaging of cell wells (IVIS imaging system operated for infrared fluorescence 
      detection). CONCLUSIONS: Despite a propensity for non-specific binding when used 
      at high concentrations and limited sensitivity, Cy7-conjugated 
      peptidase-resistant B(2)R ligands support original imaging and cytofluorometric 
      applications.
FAU - Gera, Lajos
AU  - Gera L
AD  - Department of Biochemistry, University of Colorado Denver, Aurora, CO, 80045, 
      USA.
FAU - Charest-Morin, Xavier
AU  - Charest-Morin X
AD  - Axe maladies infectieuses et immunitaires, Centre de recherche, CHU de Quebec 
      Universite Laval, Quebec, QC, G1V 4G2, Canada.
FAU - Jean, Melissa
AU  - Jean M
AD  - Axe endocrinologie et nephrologie, Centre de recherche, CHU de Quebec Universite 
      Laval, Quebec, QC, G1V 4G2, Canada.
FAU - Bachelard, Helene
AU  - Bachelard H
AD  - Axe endocrinologie et nephrologie, Centre de recherche, CHU de Quebec Universite 
      Laval, Quebec, QC, G1V 4G2, Canada.
FAU - Marceau, Francois
AU  - Marceau F
AUID- ORCID: 0000-0003-1691-6083
AD  - Centre de Recherche du CHU de Quebec (CHUL), Room T1-49, 2705 Boulevard Laurier, 
      Quebec City, QC, G1V 4G2, Canada. francois.marceau@crchul.ulaval.ca.
LA  - eng
PT  - Journal Article
DEP - 20160926
PL  - England
TA  - BMC Res Notes
JT  - BMC research notes
JID - 101462768
PMC - PMC5037861
OTO - NOTNLM
OT  - Bradykinin B2 receptors
OT  - Cyanine dye 7
OT  - Cytofluorometry
OT  - Fluorescence
OT  - Human umbilical vein
OT  - Microscopy
EDAT- 2016/09/28 06:00
MHDA- 2016/09/28 06:00
PMCR- 2016/09/26
CRDT- 2016/09/28 06:00
PHST- 2016/05/07 00:00 [received]
PHST- 2016/09/20 00:00 [accepted]
PHST- 2016/09/28 06:00 [entrez]
PHST- 2016/09/28 06:00 [pubmed]
PHST- 2016/09/28 06:00 [medline]
PHST- 2016/09/26 00:00 [pmc-release]
AID - 10.1186/s13104-016-2258-1 [pii]
AID - 2258 [pii]
AID - 10.1186/s13104-016-2258-1 [doi]
PST - epublish
SO  - BMC Res Notes. 2016 Sep 26;9(1):452. doi: 10.1186/s13104-016-2258-1.