TY - JOUR
T1 - Screening for interacting proteins with peptide biomarker of blood-brain barrier alteration under inflammatory conditions
T2 - Tamizaje para proteínas que interactúan con péptido biomarcador de alteraciones en barrera hematoencefálica bajo condiciones de neuroinflamación
AU - Vargas-Sánchez, Karina
AU - Losada Barragán, Monica
AU - Mogilevskaya, Maria
AU - Novoa-Herrán, Susana
AU - Medina Medina, Yehidi
AU - Buendía, Cristian Ignacio
AU - Lorett-Velásquez, Vaneza
AU - Martínez Bernal, Jessica
AU - Gonzalez Reyes, Rodrigo Esteban
AU - Ramírez, David
AU - Petry, Klaus G.
PY - 2021/4/29
Y1 - 2021/4/29
N2 - Neurodegenerative diseases are characterized by increased permeability of the blood–brain bar-rier (BBB) due to alterations in cellular and structural components of the neurovascular unit, par-ticularly in association with neuroinflammation. A previous screening study of peptide ligands to identify molecular alterations of the BBB in neuroinflammation by phage-display, revealed that phage clone 88 presented specific binding affinity to endothelial cells under inflammatory conditions in vivo and in vitro. Here, we aimed to identify the possible target receptor of the peptide ligand 88 expressed under inflammatory conditions. A cross-link test between phage-peptide-88 with IL-1β-stimulated human hCMEC cells, followed by mass spectrometry analysis, was used to identify the target of peptide-88. We modeled the epitope–receptor molecular inter-action between peptide-88 and its target by using docking simulations. Three proteins were se-lected as potential target candidates and tested in enzyme-linked immunosorbent assays with peptide-88: fibronectin, laminin subunit α5 and laminin subunit β-1. Among them, only laminin subunit β-1 presented measurable interaction with peptide-88. Peptide-88 showed specific inter-action with laminin subunit β-1, highlighting its importance as a potential biomarker of the laminin changes that may occur at the BBB endothelial cells under pathological inflammation conditions
AB - Neurodegenerative diseases are characterized by increased permeability of the blood–brain bar-rier (BBB) due to alterations in cellular and structural components of the neurovascular unit, par-ticularly in association with neuroinflammation. A previous screening study of peptide ligands to identify molecular alterations of the BBB in neuroinflammation by phage-display, revealed that phage clone 88 presented specific binding affinity to endothelial cells under inflammatory conditions in vivo and in vitro. Here, we aimed to identify the possible target receptor of the peptide ligand 88 expressed under inflammatory conditions. A cross-link test between phage-peptide-88 with IL-1β-stimulated human hCMEC cells, followed by mass spectrometry analysis, was used to identify the target of peptide-88. We modeled the epitope–receptor molecular inter-action between peptide-88 and its target by using docking simulations. Three proteins were se-lected as potential target candidates and tested in enzyme-linked immunosorbent assays with peptide-88: fibronectin, laminin subunit α5 and laminin subunit β-1. Among them, only laminin subunit β-1 presented measurable interaction with peptide-88. Peptide-88 showed specific inter-action with laminin subunit β-1, highlighting its importance as a potential biomarker of the laminin changes that may occur at the BBB endothelial cells under pathological inflammation conditions
U2 - 10.3390/ijms22094725
DO - 10.3390/ijms22094725
M3 - Article
C2 - 33946948
SN - 1661-6596
VL - 22
SP - 1
EP - 21
JO - International Journal of Molecular Sciences
JF - International Journal of Molecular Sciences
IS - 9
M1 - 4725
ER -