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PMID: 1377696 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Structure of the axonal surface recognition molecule neurofascin and its relationship to a neural subgroup of the immunoglobulin superfamily.

The Journal of cell biology ·Vol. 118 ·No. 1 ·1992-07-00 ·Pages 149-61

Volkmer H, Hassel B, Wolff JM, Frank R, Rathjen FG

Abstract

The chick axon-associated surface glycoprotein neurofascin is implicated in axonal growth and fasciculation as revealed by antibody perturbation experiments. Here we report the complete cDNA sequence of neurofascin. It is composed of four structural elements: At the NH2 terminus neurofascin contains six Ig-like motifs of the C2 subcategory followed by four fibronectin type III (FNIII)-related repeats. Between the FNIII-like repeats and the plasma membrane spanning region neurofascin contains a domain 75-amino acid residues-long rich in proline, alanine and threonine which might be the target of extensive O-linked glycosylation. A transmembrane segment is followed by a 113-amino acid residues-long cytoplasmic domain. Sequence comparisons indicate that neurofascin is most closely related to chick Nr-CAM and forms with L1 (Ng-CAM) and Nr-CAM a subgroup within the vertebrate Ig superfamily. Sequencing of several overlapping cDNA probes reveals interesting heterogeneities throughout the neurofascin polypeptide. Genomic Southern blots analyzed with neurofascin cDNA clones suggest that neurofascin is encoded by a single gene and its pre-mRNA might be therefore alternatively spliced. Northern blot analysis with domain specific probes showed that neurofascin mRNAs of about 8.5 kb are expressed throughout development in embryonic brain but not in liver. Isolation of neurofascin by immunoaffinity chromatography results in several molecular mass components. To analyze their origin the amino-terminal sequences of several neurofascin components were determined. The NH2-terminal sequences of the 185, 160, and 110-135 kD components are all the same as the NH2 termini predicted by the cDNA sequence, whereas the other neurofascin components start with a sequence found in a putative alternatively spliced segment between the Ig- and FNIII-like part indicating that they are derived by proteolytic cleavage. A combination of enzymatic and chemical deglycosylation procedures and the analysis of peanut lectin binding reveals O- and N-linked carbohydrates on neurofascin components which might generate additional heterogeneity.

MeSH Terms
3T3 Cells Amino Acid Sequence Animals Axons/chemistry,metabolism Base Sequence Brain/embryology Cell Adhesion Molecules Cell Adhesion Molecules, Neuronal/genetics Cloning, Molecular Extracellular Matrix Proteins/genetics Fibronectins/genetics Glycoproteins/biosynthesis,genetics,ultrastructure Immunoglobulins/genetics Membrane Glycoproteins/biosynthesis,genetics,ultrastructure Mice Molecular Sequence Data Multigene Family/genetics Nerve Growth Factors/biosynthesis,genetics,ultrastructure Protein Conformation Sequence Homology, Nucleic Acid Tenascin Transcription, Genetic Transfection
Chemicals
Cell Adhesion Molecules Cell Adhesion Molecules, Neuronal Extracellular Matrix Proteins Fibronectins Glycoproteins Immunoglobulins Membrane Glycoproteins Nerve Growth Factors Nfasc protein, mouse Nrcam protein, mouse Tenascin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Volkmer H
Zentrum für Molekulare Neurobiologie, Hamburg, Germany.
Hassel B
Wolff J M
Frank R
Rathjen F G
References (83)
83 references, click to expand
  1. Fast and sensitive detection of protein and DNA bands by treatment with potassium permanganate.
    J Biochem Biophys Methods. 1985 May;11(1):13-20 PMID: 2409124
  2. Cloning and characterization of cDNAs encoding the complete sequence of decay-accelerating factor of human complement.
    Proc Natl Acad Sci U S A. 1987 Apr;84(7):2007-11 PMID: 2436222
  3. The mouse neuronal cell surface protein F3: a phosphatidylinositol-anchored member of the immunoglobulin superfamily related to chicken contactin.
    J Cell Biol. 1989 Aug;109(2):775-88 PMID: 2474555
  4. Neural cell adhesion molecules influence second messenger systems.
    Neuron. 1989 Jul;3(1):13-20 PMID: 2559759
  5. Tissue specific O-linked glycosylation of the neural cell adhesion molecule (N-CAM).
    Development. 1989 Apr;105(4):803-11 PMID: 2598815
  6. Distribution and possible interactions of actin-associated proteins and cell adhesion molecules of nerve growth cones.
    Development. 1989 Mar;105(3):505-19 PMID: 2612362
  7. Identification of a heparin binding domain of the neural cell adhesion molecule N-CAM using synthetic peptides.
    Neuron. 1989 Feb;2(2):1157-65 PMID: 2624745
  8. Neural cell recognition molecule F11: homology with fibronectin type III and immunoglobulin type C domains.
    Neuron. 1989 Apr;2(4):1351-61 PMID: 2627374
  9. L1-mediated axon outgrowth occurs via a homophilic binding mechanism.
    Neuron. 1989 Jun;2(6):1597-603 PMID: 2627381
  10. A protein kinase activity is associated with and specifically phosphorylates the neural cell adhesion molecule L1.
    J Neurochem. 1989 Nov;53(5):1471-8 PMID: 2677246
  11. Neural cell recognition molecule F11: membrane interaction by covalently attached phosphatidylinositol.
    Biochem Biophys Res Commun. 1989 Jun 15;161(2):931-8 PMID: 2735929
  12. The cytoplasmic domain of the cell adhesion molecule uvomorulin associates with three independent proteins structurally related in different species.
    EMBO J. 1989 Jun;8(6):1711-7 PMID: 2788574
  13. Drosophila neuroglian: a member of the immunoglobulin superfamily with extensive homology to the vertebrate neural adhesion molecule L1.
    Cell. 1989 Nov 3;59(3):447-60 PMID: 2805067
  14. Human muscle neural cell adhesion molecule (N-CAM): identification of a muscle-specific sequence in the extracellular domain.
    Cell. 1987 Sep 25;50(7):1119-30 PMID: 2887295
  15. Sequence of contactin, a 130-kD glycoprotein concentrated in areas of interneuronal contact, defines a new member of the immunoglobulin supergene family in the nervous system.
    J Cell Biol. 1988 Oct;107(4):1561-73 PMID: 3049624
  16. Axon guidance and the patterning of neuronal projections in vertebrates.
    Science. 1988 Nov 4;242(4879):692-9 PMID: 3055291
  17. Distinct roles for adhesion molecules during innervation of embryonic chick muscle.
    Dev Biol. 1988 Dec;130(2):645-70 PMID: 3058545
  18. Cell binding function of E-cadherin is regulated by the cytoplasmic domain.
    EMBO J. 1988 Dec 1;7(12):3679-84 PMID: 3061804
  19. Growth cone guidance in insects: fasciclin II is a member of the immunoglobulin superfamily.
    Science. 1988 Nov 4;242(4879):700-8 PMID: 3187519
  20. Spatial regulation of axonal glycoprotein expression on subsets of embryonic spinal neurons.
    Neuron. 1988 Apr;1(2):105-16 PMID: 3272160
  21. The neural cell adhesion molecule (NCAM) as a regulator of cell-cell interactions.
    Science. 1988 Apr 1;240(4848):53-7 PMID: 3281256
  22. The immunoglobulin superfamily--domains for cell surface recognition.
    Annu Rev Immunol. 1988;6:381-405 PMID: 3289571
  23. Cloning of the alpha chain of human platelet glycoprotein Ib: a transmembrane protein with homology to leucine-rich alpha 2-glycoprotein.
    Proc Natl Acad Sci U S A. 1987 Aug;84(16):5615-9 PMID: 3303030
  24. The 180-kD component of the neural cell adhesion molecule N-CAM is involved in cell-cell contacts and cytoskeleton-membrane interactions.
    Cell Tissue Res. 1987 Oct;250(1):227-36 PMID: 3308110
  25. Neural adhesion molecule L1 as a member of the immunoglobulin superfamily with binding domains similar to fibronectin.
    Nature. 1988 Aug 25;334(6184):701-3 PMID: 3412448
  26. Neurite outgrowth patterns in cerebellar microexplant cultures are affected by antibodies to the cell surface glycoprotein L1.
    J Neurosci. 1986 Feb;6(2):605-12 PMID: 3512794
  27. Differential contributions of Ng-CAM and N-CAM to cell adhesion in different neural regions.
    J Cell Biol. 1986 Jul;103(1):145-58 PMID: 3522601
  28. Neural cell adhesion molecule: structure, immunoglobulin-like domains, cell surface modulation, and alternative RNA splicing.
    Science. 1987 May 15;236(4803):799-806 PMID: 3576199
  29. Neurofascin: a novel chick cell-surface glycoprotein involved in neurite-neurite interactions.
    Cell. 1987 Dec 4;51(5):841-9 PMID: 3677174
  30. A new method for predicting signal sequence cleavage sites.
    Nucleic Acids Res. 1986 Jun 11;14(11):4683-90 PMID: 3714490
  31. Membrane glycoproteins involved in neurite fasciculation.
    J Cell Biol. 1987 Feb;104(2):343-53 PMID: 3805123
  32. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  33. Deglycosylation of glycoproteins by trifluoromethanesulfonic acid.
    Anal Biochem. 1981 Nov 15;118(1):131-7 PMID: 6175244
  34. Domain map of the LDL receptor: sequence homology with the epidermal growth factor precursor.
    Cell. 1984 Jun;37(2):577-85 PMID: 6327078
  35. Immunocytological and biochemical characterization of a new neuronal cell surface component (L1 antigen) which is involved in cell adhesion.
    EMBO J. 1984 Jan;3(1):1-10 PMID: 6368220
  36. Heterotypic binding between neuronal membrane vesicles and glial cells is mediated by a specific cell adhesion molecule.
    J Cell Biol. 1984 May;98(5):1746-56 PMID: 6725397
  37. Isolation of microgram quantities of proteins from polyacrylamide gels for amino acid sequence analysis.
    Methods Enzymol. 1983;91:227-36 PMID: 6855576
  38. Role of sialic acid in the mobility of membrane proteins containing O-linked oligosaccharides on polyacrylamide gel electrophoresis in sodium dodecyl sulfate.
    Eur J Biochem. 1982 Mar 1;122(3):581-6 PMID: 7060593
  39. Topologically restricted appearance in the developing chick retinotectal system of Bravo, a neural surface protein: experimental modulation by environmental cues.
    J Cell Biol. 1990 Dec;111(6 Pt 2):3087-96 PMID: 2269667
  40. The neural cell adhesion molecule N-CAM enhances L1-dependent cell-cell interactions.
    J Cell Biol. 1990 Jan;110(1):193-208 PMID: 2295682
  41. The axonal glycoprotein TAG-1 is an immunoglobulin superfamily member with neurite outgrowth-promoting activity.
    Cell. 1990 Apr 6;61(1):157-70 PMID: 2317872
  42. An L1-like molecule, the 8D9 antigen, is a potent substrate for neurite extension.
    Proc Natl Acad Sci U S A. 1987 Nov;84(21):7753-7 PMID: 3478724
  43. Extension of neurites on axons is impaired by antibodies against specific neural cell surface glycoproteins.
    J Cell Biol. 1987 Feb;104(2):355-62 PMID: 3543026
  44. Biochemical characterization of polypeptide components involved in neurite fasciculation and elongation.
    Eur J Biochem. 1987 Nov 2;168(3):551-61 PMID: 3665934
  45. The appearance of an L1-like molecule in the chick primary visual pathway.
    J Neurosci. 1986 Oct;6(10):2987-94 PMID: 3760944
  46. Involvement of the nerve growth factor-inducible large external glycoprotein (NILE) in neurite fasciculation in primary cultures of rat brain.
    Proc Natl Acad Sci U S A. 1985 Feb;82(4):1276-80 PMID: 3856258
  47. The human LDL receptor: a cysteine-rich protein with multiple Alu sequences in its mRNA.
    Cell. 1984 Nov;39(1):27-38 PMID: 6091915
  48. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum.
    Anal Biochem. 1984 Feb;137(1):266-7 PMID: 6329026
  49. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs.
    Nucleic Acids Res. 1984 Jan 25;12(2):857-72 PMID: 6694911
  50. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells.
    Mol Cell Biol. 1982 Sep;2(9):1044-51 PMID: 6960240
  51. A simple method for displaying the hydropathic character of a protein.
    J Mol Biol. 1982 May 5;157(1):105-32 PMID: 7108955
  52. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  53. A simplification of the protein assay method of Lowry et al. which is more generally applicable.
    Anal Biochem. 1977 Dec;83(2):346-56 PMID: 603028
  54. The axonally secreted cell adhesion molecule, axonin-1. Primary structure, immunoglobulin-like and fibronectin-type-III-like domains and glycosyl-phosphatidylinositol anchorage.
    Eur J Biochem. 1992 Mar 1;204(2):453-63 PMID: 1311675
  55. Binding of the integrin Mac-1 (CD11b/CD18) to the third immunoglobulin-like domain of ICAM-1 (CD54) and its regulation by glycosylation.
    Cell. 1991 Jun 14;65(6):961-71 PMID: 1675157
  56. Morphoregulatory activities of NCAM and N-cadherin can be accounted for by G protein-dependent activation of L- and N-type neuronal Ca2+ channels.
    Cell. 1991 Oct 4;67(1):21-33 PMID: 1680564
  57. Molecular mechanisms of axon growth and guidance.
    Annu Rev Cell Biol. 1991;7:117-59 PMID: 1687312
  58. Differential splicing generates a nervous system-specific form of Drosophila neuroglian.
    Neuron. 1990 May;4(5):697-709 PMID: 1693086
  59. Structure of the chicken neuron-glia cell adhesion molecule, Ng-CAM: origin of the polypeptides and relation to the Ig superfamily.
    J Cell Biol. 1991 Mar;112(5):1017-29 PMID: 1705558
  60. Neurite outgrowth on immobilized axonin-1 is mediated by a heterophilic interaction with L1(G4).
    J Cell Biol. 1991 Nov;115(4):1113-26 PMID: 1720120
  61. A comprehensive analysis of the developmental and tissue-specific expression of the isoactin multigene family in the rat.
    Dev Biol. 1991 Dec;148(2):442-58 PMID: 1743394
  62. Restrictin: a chick neural extracellular matrix protein involved in cell attachment co-purifies with the cell recognition molecule F11.
    Development. 1991 Sep;113(1):151-64 PMID: 1764992
  63. Molecular structure and functional testing of human L1CAM: an interspecies comparison.
    Genomics. 1991 Oct;11(2):416-23 PMID: 1769655
  64. Molecular cloning of NILE glycoprotein and evidence for its continued expression in mature rat CNS.
    J Neurosci Res. 1991 Nov;30(3):567-81 PMID: 1800773
  65. Cell and substrate adhesion molecules in Drosophila.
    Annu Rev Cell Biol. 1991;7:505-57 PMID: 1809354
  66. Neural cell contact and axonal growth.
    Curr Opin Cell Biol. 1991 Dec;3(6):992-1000 PMID: 1814371
  67. Extracellular matrix molecules and their receptors: functions in neural development.
    Annu Rev Neurosci. 1991;14:531-70 PMID: 1851608
  68. DM-GRASP, a novel immunoglobulin superfamily axonal surface protein that supports neurite extension.
    Neuron. 1991 Aug;7(2):209-20 PMID: 1873027
  69. Molecular cloning of cDNA encoding the rat neural cell adhesion molecule L1. Two L1 isoforms in the cytoplasmic region are produced by differential splicing.
    FEBS Lett. 1991 Sep 2;289(1):91-5 PMID: 1894011
  70. Molecular cloning and expression of a novel adhesion molecule, SC1.
    Neuron. 1991 Oct;7(4):535-45 PMID: 1931049
  71. The arrangement of the immunoglobulin-like domains of ICAM-1 and the binding sites for LFA-1 and rhinovirus.
    Cell. 1990 Apr 20;61(2):243-54 PMID: 1970514
  72. The axonally secreted protein axonin-1 is a potent substratum for neurite growth.
    J Cell Biol. 1991 Feb;112(3):449-55 PMID: 1991792
  73. Isolation and sequence of partial cDNA clones of human L1: homology of human and rodent L1 in the cytoplasmic region.
    J Neurochem. 1991 Mar;56(3):797-804 PMID: 1993895
  74. Cadherin cell adhesion receptors as a morphogenetic regulator.
    Science. 1991 Mar 22;251(5000):1451-5 PMID: 2006419
  75. Structure of a new nervous system glycoprotein, Nr-CAM, and its relationship to subgroups of neural cell adhesion molecules.
    J Cell Biol. 1991 Jun;113(6):1399-412 PMID: 2045418
  76. Guidance of developing axons by diffusible chemoattractants.
    Cold Spring Harb Symp Quant Biol. 1990;55:279-89 PMID: 2132821
  77. Place-dependent cell adhesion, process retraction, and spatial signaling in neural morphogenesis.
    Cold Spring Harb Symp Quant Biol. 1990;55:303-18 PMID: 2132823
  78. Molecular genetics of neuronal recognition in Drosophila: evolution and function of immunoglobulin superfamily cell adhesion molecules.
    Cold Spring Harb Symp Quant Biol. 1990;55:327-40 PMID: 2132825
  79. Neurite outgrowth in response to transfected N-CAM changes during development and is modulated by polysialic acid.
    Neuron. 1990 Aug;5(2):209-19 PMID: 2200449
  80. Why are proteins O-glycosylated?
    Trends Biochem Sci. 1990 Aug;15(8):291-4 PMID: 2204153
  81. The avian tectobulbar tract: development, explant culture, and effects of antibodies on the pattern of neurite outgrowth.
    J Neurosci. 1990 Sep;10(9):3118-34 PMID: 2204687
  82. Protein kinase recognition sequence motifs.
    Trends Biochem Sci. 1990 Sep;15(9):342-6 PMID: 2238044
  83. Neurite outgrowth promoting activity of G4 and its inhibition by monoclonal antibodies.
    J Neurosci Res. 1990 Feb;25(2):180-6 PMID: 2319627
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.118.1.149
Published
1992-07-00
Pages
149-61
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2289533
Subset
IM
Databases
GENBANK
M31165, M31166, M74319, M74320, M74321, M74322, M74323, M74324, X65191, X65224
External Links
PubMed source DOI: 10.1083/jcb.118.1.149 PMC Full Text View journal details
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