TY - JOUR
T1 - Increasing paracellular porosity by E-cadherin peptides
T2 - Discovery of bulge and groove regions in the EC1-domain of E-cadherin
AU - Sinaga, Ernawati
AU - Jois, Seetharama D.S.
AU - Avery, Mike
AU - Makagiansar, Irwan T.
AU - Tambunan, Usman S.F.
AU - Audus, Kenneth L.
AU - Siahaan, Teruna J.
PY - 2002/8
Y1 - 2002/8
N2 - Purpose. The objective of this work is to evaluate the ability of peptides derived from the bulge (HAV-peptides) and groove (ADT-peptides) regions of E-cadherin EC1-domain to increase the paracellular porosity of the intercellular junctions of Madin-Darby canine kidney (MDCK) cell monolayers. Methods. Peptides were synthesized using a solid-phase method and were purified using semi-preparative HPLC. MDCK monolayers were used to evaluate the ability of cadherin peptides to modulate cadherin-cadherin interactions in the intercellular junctions. The increase in intercellular junction porosity was determined by the change in transepithelial electrical resistance (TEER) values and the paracellular transport of 14C-mannitol. Results. HAV- and ADT-peptides can lower the TEER value of MDCK cell monolayers and enhance the paracellular permeation of 14C-mannitol. HAV- and ADT-decapeptides can modulate the intercellular junctions when they are added from the basolateral side but not from the apical side; on the other hand, HAV- and ADT-hexapeptides increase the paracellular porosity of the monolayers when added from either side. Conjugation of HAV- and ADT-peptides using ω-aminocaproic acid can only work to modulate the paracellular porosity when ADT-peptide is at the N-terminus and HAV-peptide is at the C-terminus; because of its size, the conjugate can only modulate the intercellular junction when added from the basolateral side. Conclusions. Peptides from the bulge and groove regions of the ECl domain of E-cadherin can inhibit cadherin-cadherin interactions, resulting in the opening of the paracellular junctions. These peptides may be used to improve paracellular permeation of peptides and proteins. Furthermore, this work suggests that both groove and bulge regions of EC-domain are important for cadherin-cadherin interactions.
AB - Purpose. The objective of this work is to evaluate the ability of peptides derived from the bulge (HAV-peptides) and groove (ADT-peptides) regions of E-cadherin EC1-domain to increase the paracellular porosity of the intercellular junctions of Madin-Darby canine kidney (MDCK) cell monolayers. Methods. Peptides were synthesized using a solid-phase method and were purified using semi-preparative HPLC. MDCK monolayers were used to evaluate the ability of cadherin peptides to modulate cadherin-cadherin interactions in the intercellular junctions. The increase in intercellular junction porosity was determined by the change in transepithelial electrical resistance (TEER) values and the paracellular transport of 14C-mannitol. Results. HAV- and ADT-peptides can lower the TEER value of MDCK cell monolayers and enhance the paracellular permeation of 14C-mannitol. HAV- and ADT-decapeptides can modulate the intercellular junctions when they are added from the basolateral side but not from the apical side; on the other hand, HAV- and ADT-hexapeptides increase the paracellular porosity of the monolayers when added from either side. Conjugation of HAV- and ADT-peptides using ω-aminocaproic acid can only work to modulate the paracellular porosity when ADT-peptide is at the N-terminus and HAV-peptide is at the C-terminus; because of its size, the conjugate can only modulate the intercellular junction when added from the basolateral side. Conclusions. Peptides from the bulge and groove regions of the ECl domain of E-cadherin can inhibit cadherin-cadherin interactions, resulting in the opening of the paracellular junctions. These peptides may be used to improve paracellular permeation of peptides and proteins. Furthermore, this work suggests that both groove and bulge regions of EC-domain are important for cadherin-cadherin interactions.
KW - ADT peptides
KW - Adherens junction
KW - Cell-cell adhesion
KW - E-cadherin
KW - HAV peptides
KW - Intercellular junctions
KW - MDCK cell monolayers
UR - http://www.scopus.com/inward/record.url?scp=0036692383&partnerID=8YFLogxK
U2 - 10.1023/A:1019850226631
DO - 10.1023/A:1019850226631
M3 - Article
C2 - 12240943
AN - SCOPUS:0036692383
SN - 0724-8741
VL - 19
SP - 1170
EP - 1179
JO - Pharmaceutical Research
JF - Pharmaceutical Research
IS - 8
ER -