Brain-specific angiogenesis inhibitor 1 is expressed in the Myo/Nog cell lineage.
Amino Acid Substitution
Angiogenic Proteins
/ biosynthesis
Animals
Antibodies, Monoclonal
/ immunology
Antibody Specificity
Antigen-Antibody Reactions
Brain
/ cytology
Carrier Proteins
/ analysis
Cell Lineage
Epitopes
/ immunology
Eye Proteins
/ biosynthesis
Humans
Male
Mice
Mice, Inbred C57BL
Models, Molecular
Muscle Development
MyoD Protein
/ analysis
Myofibroblasts
/ metabolism
Organ Specificity
Protein Conformation
Protein Domains
Rabbits
Rats, Sprague-Dawley
Receptors, G-Protein-Coupled
/ biosynthesis
Repetitive Sequences, Amino Acid
Skin
/ cytology
Species Specificity
Tattooing
Young Adult
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2020
2020
Historique:
received:
15
03
2020
accepted:
02
06
2020
entrez:
3
7
2020
pubmed:
3
7
2020
medline:
9
9
2020
Statut:
epublish
Résumé
The Myo/Nog cell lineage was discovered in the chick embryo and is also present in adult mammalian tissues. The cells are named for their expression of mRNA for the skeletal muscle specific transcription factor MyoD and bone morphogenetic protein inhibitor Noggin. A third marker for Myo/Nog cells is the cell surface molecule recognized by the G8 monoclonal antibody (mAb). G8 has been used to detect, track, isolate and kill Myo/Nog cells. In this study, we screened a membrane proteome array for the target of the G8 mAb. The array consisted of >5,000 molecules, each synthesized in their native confirmation with appropriate post-translational modifications in a single clone of HEK-293T cells. G8 mAb binding to the clone expressing brain-specific angiogenesis inhibitor 1 (BAI1) was detected by flow cytometry, re-verified by sequencing and validated by transfection with the plasmid construct for BAI1. Further validation of the G8 target was provided by enzyme-linked immunosorbent assay. The G8 epitope was identified by screening a high-throughput, site directed mutagenesis library designed to cover 95-100% of the 954 amino acids of the extracellular domain of the BAI1 protein. The G8 mAb binds within the third thrombospondin repeat of the extracellular domain of human BAI1. Immunofluorescence localization experiments revealed that G8 and a commercially available BAI1 mAb co-localize to the subpopulation of Myo/Nog cells in the skin, eyes and brain. Expression of the multi-functional BAI1 protein in Myo/Nog cells introduces new possibilities for the roles of Myo/Nog cells in normal and diseased tissues.
Identifiants
pubmed: 32614850
doi: 10.1371/journal.pone.0234792
pii: PONE-D-20-06868
pmc: PMC7332021
doi:
Substances chimiques
ADGRB1 protein, human
0
Angiogenic Proteins
0
Antibodies, Monoclonal
0
Carrier Proteins
0
Epitopes
0
Eye Proteins
0
MyoD Protein
0
MyoD1 myogenic differentiation protein
0
Receptors, G-Protein-Coupled
0
noggin protein
148294-77-3
Types de publication
Comparative Study
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0234792Déclaration de conflit d'intérêts
This work was supported by an anonymous donation for the Myo/Nog cell program project to MG-W and AB-N. Funding was also provided by Genisphere, LLC, https://genisphere.com/. Employees of Genisphere participated in study design, data collection and analysis, the decision to publish and preparation of the manuscript. A provisional patent has been submitted describing uses of the G8 monoclonal antibody. Related products are in development. No income is derived from these products. This does not alter our adherence to PLOS ONE policies on sharing data and materials.
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