PMID- 24790958
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20140505
LR  - 20211021
IS  - 2296-2646 (Print)
IS  - 2296-2646 (Electronic)
IS  - 2296-2646 (Linking)
VI  - 1
DP  - 2013
TI  - Synthesis of fluorescent analogs of relaxin family peptides and their preliminary 
      in vitro and in vivo characterization.
PG  - 30
LID - 10.3389/fchem.2013.00030 [doi]
LID - 30
AB  - Relaxin, a heterodimeric polypeptide hormone, is a key regulator of collagen 
      metabolism and multiple vascular control pathways in humans and rodents. Its 
      actions are mediated via its cognate G-protein-coupled receptor, RXFP1 although 
      it also "pharmacologically" activates RXFP2, the receptor for the related, 
      insulin-like peptide 3 (INSL3), which has specific actions on reproduction and 
      bone metabolism. Therefore, experimental tools to facilitate insights into the 
      distinct biological actions of relaxin and INSL3 are required, particularly for 
      studies of tissues containing both RXFP1 and RXFP2. Here, we chemically 
      functionalized human (H2) relaxin, the RXFP1-selective relaxin analog 
      H2:A(4-24)(F23A), and INSL3 to accommodate a fluorophore without marked reduction 
      in binding or activation propensity. Chemical synthesis of the two chains for 
      each peptide was followed by sequential regioselective formation of their three 
      disulfide bonds. Click chemistry conjugation of Cy5.5 at the B-chain N-terminus, 
      with conservation of the disulfide bonds, yielded analogs displaying appropriate 
      selective binding affinity and ability to activate RXFP1 and/or RXFP2 in vitro. 
      The in vivo biological activity of Cy5.5-H2 relaxin and Cy5.5-H2:A(4-24)(F23A) 
      was confirmed in mice, as acute intracerebroventricular (icv) infusion of these 
      peptides (but not Cy5.5-INSL3) stimulated water drinking, an established 
      behavioral response elicited by central RXFP1 activation. The central 
      distribution of Cy5.5-conjugated peptides was examined in mice killed 30 min 
      after infusion, revealing higher fluorescence within brain tissue near-adjacent 
      to the cerebral ventricle walls relative to deeper brain areas. Production of 
      fluorophore-conjugated relaxin family peptides will facilitate future 
      pharmacological studies to probe the function of H2 relaxin/RXFP1 and INSL3/RXFP2 
      signaling in vivo while tracking their distribution following central or 
      peripheral administration.
FAU - Chan, Linda J
AU  - Chan LJ
AD  - The Florey Institute of Neuroscience and Mental Health, The University of 
      Melbourne VIC, Australia ; School of Chemistry, The University of Melbourne VIC, 
      Australia.
FAU - Smith, Craig M
AU  - Smith CM
AD  - The Florey Institute of Neuroscience and Mental Health, The University of 
      Melbourne VIC, Australia ; Florey Department of Neuroscience and Mental Health, 
      The University of Melbourne VIC, Australia.
FAU - Chua, Berenice E
AU  - Chua BE
AD  - The Florey Institute of Neuroscience and Mental Health, The University of 
      Melbourne VIC, Australia.
FAU - Lin, Feng
AU  - Lin F
AD  - The Florey Institute of Neuroscience and Mental Health, The University of 
      Melbourne VIC, Australia.
FAU - Bathgate, Ross A D
AU  - Bathgate RA
AD  - The Florey Institute of Neuroscience and Mental Health, The University of 
      Melbourne VIC, Australia ; Florey Department of Neuroscience and Mental Health, 
      The University of Melbourne VIC, Australia ; Department of Biochemistry and 
      Molecular Biology, The University of Melbourne VIC, Australia.
FAU - Separovic, Frances
AU  - Separovic F
AD  - School of Chemistry, The University of Melbourne VIC, Australia.
FAU - Gundlach, Andrew L
AU  - Gundlach AL
AD  - The Florey Institute of Neuroscience and Mental Health, The University of 
      Melbourne VIC, Australia ; Florey Department of Neuroscience and Mental Health, 
      The University of Melbourne VIC, Australia.
FAU - Hossain, Mohammed Akhter
AU  - Hossain MA
AD  - The Florey Institute of Neuroscience and Mental Health, The University of 
      Melbourne VIC, Australia ; School of Chemistry, The University of Melbourne VIC, 
      Australia ; Florey Department of Neuroscience and Mental Health, The University 
      of Melbourne VIC, Australia.
FAU - Wade, John D
AU  - Wade JD
AD  - The Florey Institute of Neuroscience and Mental Health, The University of 
      Melbourne VIC, Australia ; School of Chemistry, The University of Melbourne VIC, 
      Australia ; Florey Department of Neuroscience and Mental Health, The University 
      of Melbourne VIC, Australia.
LA  - eng
PT  - Journal Article
DEP - 20131206
PL  - Switzerland
TA  - Front Chem
JT  - Frontiers in chemistry
JID - 101627988
PMC - PMC3982560
OTO - NOTNLM
OT  - Cy5.5 fluorophore
OT  - RXFP1
OT  - RXFP2
OT  - brain
OT  - click chemistry
OT  - relaxin
EDAT- 2013/01/01 00:00
MHDA- 2013/01/01 00:01
PMCR- 2013/01/01
CRDT- 2014/05/03 06:00
PHST- 2013/10/08 00:00 [received]
PHST- 2013/11/18 00:00 [accepted]
PHST- 2014/05/03 06:00 [entrez]
PHST- 2013/01/01 00:00 [pubmed]
PHST- 2013/01/01 00:01 [medline]
PHST- 2013/01/01 00:00 [pmc-release]
AID - 10.3389/fchem.2013.00030 [doi]
PST - epublish
SO  - Front Chem. 2013 Dec 6;1:30. doi: 10.3389/fchem.2013.00030. eCollection 2013.