Mutational Analysis of the L1 Neuronal Cell Adhesion Molecule Identifies Membrane-Proximal Amino Acids of the Cytoplasmic Domain That Are Required for Cytoskeletal Anchorage

Dahlin-Huppe, Kimberlee; Berglund, Erik; Ranscht, Barbara; Stallcup, William B.

doi:10.1006/mcne.1997.0608

Cited by 55 publications

(40 citation statements)

References 64 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…ELISA results showed increased and dose-dependent L1-Fc increased exposure of the neuritogenesis-promoting epitope on PA containing substrates compared to PDL controls. These results support a more outward display of L1 on PA-treated substrates, given the nature and location of antibody binding epitopes within the N-terminal, first and second Ig-like domains of the ectodomain of L1 [43]. The amount of L1-Fc adsorbed on the polymer thin films was quantified using QCM-D. Representative QCM-D plots of frequency over time show subsequent frequency drops indicating deposition of PDL, PA, and L1-Fc, respectively (Fig.…”

Section: Protein a Coated Polymer Films Support Increased Surface Adssupporting

confidence: 69%

“…Our studies with cyclic RGD peptide pretreatment showed reduction in SCN attachment and neurite outgrowth when plated onto L1-Fc/ PA/PDL and L1-Fc/PA treated thin films, suggesting that this domain was also exposed and available for a v b 3 integrin binding. Thus, we propose that the PA-based treatment of the polymers not only facilitated oriented display of L1-Fc, but also engendered a display of L1-Fc molecules in close proximity to one another, which may mirror the close apposition of highly organized and clustered L1 molecules on the cell surface [43,64]. Using AFM imaging of substrate counterstained with L1 antibodies, we established that L1-Fc/PA substrates showed adsorbed L1-Fc.…”

Section: Discussionmentioning

confidence: 83%

See 1 more Smart Citation

Oriented, Multimeric Biointerfaces of the L1 Cell Adhesion Molecule: An Approach to Enhance Neuronal and Neural Stem Cell Functions on 2-D and 3-D Polymer Substrates

et al. 2012

View full text Add to dashboard Cite

This article focuses on elucidating the key presentation features of neurotrophic ligands at polymer interfaces. Different biointerfacial configurations of the human neural cell adhesion molecule L1 were established on two-dimensional films and three-dimensional fibrous scaffolds of synthetic tyrosine-derived polycarbonate polymers and probed for surface concentrations, microscale organization, and effects on cultured primary neurons and neural stem cells. Underlying polymer substrates were modified with varying combinations of protein A and poly-d-lysine to modulate the immobilization and presentation of the Fc fusion fragment of the extracellular domain of L1 (L1-Fc). When presented as an oriented and multimeric configuration from protein A-pretreated polymers, L1-Fc significantly increased neurite outgrowth of rodent spinal cord neurons and cerebellar neurons as early as 24 h compared to the traditional presentation via adsorption onto surfaces treated with poly-d-lysine. Cultures of human neural progenitor cells screened on the L1-Fc/polymer biointerfaces showed significantly enhanced neuronal differentiation and neuritogenesis on all protein A oriented substrates. Notably, the highest degree of βIII-tubulin expression for cells in 3-D fibrous scaffolds were observed in protein A oriented substrates with PDL pretreatment, suggesting combined effects of cell attachment to polycationic charged substrates with subcellular topography along with L1-mediated adhesion mediating neuronal differentiation. Together, these findings highlight the promise of displays of multimeric neural adhesion ligands via biointerfacially engineered substrates to “cooperatively” enhance neuronal phenotypes on polymers of relevance to tissue engineering.

show abstract

Section: Protein a Coated Polymer Films Support Increased Surface Adssupporting

confidence: 69%

Section: Discussionmentioning

confidence: 83%

Oriented, Multimeric Biointerfaces of the L1 Cell Adhesion Molecule: An Approach to Enhance Neuronal and Neural Stem Cell Functions on 2-D and 3-D Polymer Substrates

et al. 2012

View full text Add to dashboard Cite

show abstract

“…We have also used two chimeric molecules containing almost the entire ectodomain of NG2 (terminating after amino acid residue Leu-2218) but lacking the NG2 transmembrane and cytoplasmic domains. In the case of the NG2/contactin chimera (NG2/CNTN), this segment of the NG2 polypeptide is replaced by the glycosylphosphatidylinositol (GPI) linkage region of human contactin (Berglund and Ranscht, 1994;Dahlin-Huppe et al, 1997). In the case of the NG2/L1 chimera, the same segment is replaced by the membrane-spanning and cytoplasmic domains of the human L1 neuronal cell adhesion molecule (Hlavin and Lemmon, 1991;Dahlin-Huppe et al, 1997).…”

Section: Cdna Constructs and Transfectionsmentioning

confidence: 99%

“…In the case of the NG2/contactin chimera (NG2/CNTN), this segment of the NG2 polypeptide is replaced by the glycosylphosphatidylinositol (GPI) linkage region of human contactin (Berglund and Ranscht, 1994;Dahlin-Huppe et al, 1997). In the case of the NG2/L1 chimera, the same segment is replaced by the membrane-spanning and cytoplasmic domains of the human L1 neuronal cell adhesion molecule (Hlavin and Lemmon, 1991;Dahlin-Huppe et al, 1997). In both cases, a segment of the NG2 cDNA coding for the transmembrane and cytoplasmic domains was excised from the pcDNA/NG2 plasmid and replaced by a PCR product coding for the desired segment of either contactin or L1.…”

Section: Cdna Constructs and Transfectionsmentioning

confidence: 99%

“…Immunoprecipitation of detergent-extracted material from 125 I-labeled cells was performed as described previously (Nishiyama et al, 1991;Dahlin-Huppe et al, 1997). In some experiments, the 125 Ilabeled cells were treated with phosphoinositol-specific phospholipase C (Oxford Glycosystems, Abingdon Oxon, United Kingdom) before detergent extraction and immunoprecipitation.…”

Section: Immunoprecipitationmentioning

confidence: 99%

See 1 more Smart Citation

Cytoskeletal Reorganization Induced by Engagement of the NG2 Proteoglycan Leads to Cell Spreading and Migration

Fang

Burg

Barritt

et al. 1999

MBoC

Self Cite

View full text Add to dashboard Cite

Cells expressing the NG2 proteoglycan can attach, spread, and migrate on surfaces coated with NG2 mAbs, demonstrating that engagement of NG2 can trigger the cytoskeletal rearrangements necessary for changes in cell morphology and motility. Engagement of different epitopes of the proteoglycan results in distinct forms of actin reorganization. On mAb D120, the cells contain radial actin spikes characteristic of filopodial extension, whereas on mAb N143, the cells contain cortical actin bundles characteristic of lamellipodia. Cells that express NG2 variants lacking the transmembrane and cytoplasmic domains are unable to spread or migrate on NG2 mAb-coated surfaces, indicating that these portions of the molecule are essential for NG2-mediated signal transduction. Cells expressing an NG2 variant lacking the C-terminal half of the cytoplasmic domain can still spread normally on mAbs D120 and N143, suggesting that the membraneproximal cytoplasmic segment is responsible for this process. In contrast, this variant migrates poorly on mAb D120 and exhibits abnormal arrays of radial actin filaments decorated with fascin during spreading on this mAb. The C-terminal portion of the NG2 cytoplasmic domain, therefore, may be involved in regulating molecular events that are crucial for cell motility.

show abstract

Cellular signaling by neural cell adhesion molecules of the immunoglobulin superfamily

2000

View full text Add to dashboard Cite

Mutational Analysis of the L1 Neuronal Cell Adhesion Molecule Identifies Membrane-Proximal Amino Acids of the Cytoplasmic Domain That Are Required for Cytoskeletal Anchorage

Cited by 55 publications

References 64 publications

Oriented, Multimeric Biointerfaces of the L1 Cell Adhesion Molecule: An Approach to Enhance Neuronal and Neural Stem Cell Functions on 2-D and 3-D Polymer Substrates

Oriented, Multimeric Biointerfaces of the L1 Cell Adhesion Molecule: An Approach to Enhance Neuronal and Neural Stem Cell Functions on 2-D and 3-D Polymer Substrates

Cytoskeletal Reorganization Induced by Engagement of the NG2 Proteoglycan Leads to Cell Spreading and Migration

Cellular signaling by neural cell adhesion molecules of the immunoglobulin superfamily

Contact Info

Product

Resources

About