Identification and validation of a Lewis x glycomimetic peptide.
Glycans play important roles in regulating cell recognition and interactions to fine tune development, and synaptic plasticity and regeneration in the adult nervous system. The spatial and temporal expression pattern of Lewis(x) (a terminal trisaccharide epitope characterized by alpha1,3-fucosyl-N-acetyl-lactosamine) in the nervous system indicates an important role of this epitope in neurogenesis and brain development. Localization of Lewis(x) in the proliferative subventricular zone of the developing nervous system and also its expression on stem cells of the adult nervous system suggests a role in neurogenesis and hence regeneration. To provide an alternative tool to elucidate the functional roles of Lewis(x), we screened a random peptide phage library against a Lewis(x)-specific antibody to identify a Lewis(x) glycomimetic peptide. We identified a peptide that specifically bound to the Lewis(x)-specific antibody and this binding could be competed by the Lewis(x) glycan. Different aspects of the Lewis(x) glycomimetic peptide were investigated by introducing it in in vitro assays measuring neurite outgrowth and in in vivo assays to determine its efficacy in regeneration of peripheral nerve and spinal cord after injury in adult mice. In vitro, neurite outgrowth triggered by the Lewis(x-)carrying adhesion molecule CD24 was abolished alike by the Lewis(x) glycan and the glycomimetic peptide, while no influence of the glycomimetic peptide was seen in regeneration. Our results validate the use of Lewis(x) glycomimetic peptide as a functionally equivalent structure to the Lewis(x) glycan.
Katagihallimath
Nainesh
N
Zentrum für Molekulare Neurobiologie, Universitätsklinikum Hamburg-Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany.
Mehanna
Ali
A
Guseva
Daria
D
Kleene
Ralf
R
Schachner
Melitta
M
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D051927:CD24 Antigen / Q000737:chemistry / Q000378:metabolism; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology; D004797:Enzyme-Linked Immunosorbent Assay; D005267:Femoral Nerve / Q000473:pathology / Q000503:physiopathology; D006020:Glycopeptides / Q000737:chemistry / Q000378:metabolism*; D016256:Lewis X Antigen / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009043:Motor Activity / Q000502:physiology; D009416:Nerve Regeneration / Q000502:physiology; D016501:Neurites / Q000378:metabolism; D019151:Peptide Library; D020127:Recovery of Function / Q000502:physiology; D013119:Spinal Cord Injuries / Q000503:physiopathology; D013997:Time Factors; D054928:Validation Studies as Topic
D051927:CD24 Antigen; D006020:Glycopeptides; D016256:Lewis X Antigen; D019151:Peptide Library