Improving the selectivity of HAV-peptides in modulating E-cadherin-E-cadherin interactions in the intercellular junction of MDCK cell monolayers.
OBJECTIVE
The objective of this work is to understand the sequence specificity of HAV peptides and to improve their selectivity in regulating E-cadherin-E-cadherin interactions in the intercellular junctions.


METHODS
Peptide 1 was modified using an alanine scanning method to give peptides 2-6. The ability of these peptides to modulate intercellular junctions was evaluated using Madin-Darby Canine Kidney (MDCK) cell monolayers on Transwell membranes from either the apical (AP) or the basolateral (BL) side. Modulation of the intercellular junctions was measured by the ability to lower the transepithelial electrical resistance (TEER) of MDCK monolayers and by the increase in mannitol flux. Molecular docking experiments were performed to model the binding properties of these peptides to the EC1 domain of E-cadherin.


RESULTS
Peptides 5 (Ac-SHAVAS-NH2) and 6 (Ac-SHAVSA-NH2) were found to be more effective than the parent peptide 1 in decreasing the resistance of the cell monolayer. Furthermore, comparative studies with the control and the weak inhibitor peptide 2 indicate that peptide 5 displayed a significant increase in mannitol flux. Molecular docking of peptides 1, 2 and 5 to the EC1 domain suggests that peptide 5 has the lowest binding energy.


CONCLUSIONS
HAV peptides have the ability to modulate E-cadherin-E-cadherin interactions in the intercellular junctions of the MDCK cell monolayer, thus indirectly increasing the permeability of the tight junctions. This observation indicates that residues flanking the HAV sequence are important in the binding selectivity of HAV peptides to E-cadherin. Molecular docking can further aid in the design of peptides with better selectivity to the EC1 domain of E-cadherin.
Makagiansar
I T
IT
Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence 66045, USA.
Avery
M
M
Hu
Y
Y
Audus
K L
KL
Siahaan
T J
TJ
D000409:Alanine / Q000494:pharmacology*; D000818:Animals; D015820:Cadherins / Q000378:metabolism*; D002450:Cell Communication / Q000187:drug effects*; D002460:Cell Line / Q000187:drug effects / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D004285:Dogs; D017097:Electric Impedance; D006639:Histidine / Q000494:pharmacology*; D007365:Intercellular Junctions / Q000187:drug effects* / Q000378:metabolism; D009842:Oligopeptides / Q000494:pharmacology*; D014633:Valine / Q000494:pharmacology*
D015820:Cadherins; D009842:Oligopeptides; D006639:Histidine; D014633:Valine; D000409:Alanine