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PMID: 4030898 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Purification of a factor that promotes neurite outgrowth: isolation of laminin and associated molecules.

The Journal of cell biology ·Vol. 101 ·No. 3 ·1985-09-00 ·Pages 898-913

Lander AD, Fujii DK, Reichardt LF

Abstract

When culture medium, conditioned by any of several cell types, is applied to a polycationic substratum, a substance is adsorbed that causes neurons cultured on that substratum to extend processes (neurites) rapidly and profusely. We have purified the factor responsible for this effect from medium conditioned by bovine corneal endothelial cells, and have shown that it is composed of the glycoprotein laminin and two associated laminin-binding molecules: a sulfated protein known as entactin, and a large heparan sulfate proteoglycan. Of these molecules, only laminin was found to be present throughout the purification in all fractions possessing neurite outgrowth-promoting activity and absent from all fractions lacking activity. Laminin, purified from other sources, has been shown previously to promote extensive outgrowth by cultured neurons. These and other data presented here support the conclusion that laminin is responsible for the neurite outgrowth-promoting activity of the conditioned medium factor. Evidence is also presented that the association of a proteoglycan with laminin promotes efficient attachment of laminin to polycationic substrata, particularly in the presence of competing molecules.

MeSH Terms
Animals Cattle Cell Adhesion Cell Differentiation Cells, Cultured Cornea/cytology Culture Media Epithelial Cells Extracellular Matrix/physiology Ganglia, Sympathetic/cytology Glycoproteins/isolation & purification,physiology Heparitin Sulfate/isolation & purification,physiology Laminin/isolation & purification,physiology Membrane Glycoproteins Neurons/cytology Proteoglycans/isolation & purification,physiology Rats
Chemicals
Culture Media Glycoproteins Laminin Membrane Glycoproteins Proteoglycans nidogen Heparitin Sulfate
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Lander A D
Fujii D K
Reichardt L F
References (61)
61 references, click to expand
  1. The reversible dissociation of the alkaline phosphatase of Escherichia coli. I. Formation and reactivation of subunits.
    J Biol Chem. 1965 Nov;240(11):4284-92 PMID: 4954367
  2. Laminin.
    Methods Enzymol. 1982;82 Pt A:831-8 PMID: 7078458
  3. Laminin, fibronectin, and collagen in synaptic and extrasynaptic portions of muscle fiber basement membrane.
    J Cell Biol. 1982 May;93(2):442-51 PMID: 7047538
  4. Neurite outgrowth and protein synthesis by PC12 cells as a function of substratum and nerve growth factor.
    J Neurosci. 1982 Aug;2(8):1157-75 PMID: 7108587
  5. Characterization of a factor that promotes neurite outgrowth: evidence linking activity to a heparan sulfate proteoglycan.
    J Cell Biol. 1982 Sep;94(3):574-85 PMID: 6215411
  6. Modulation of NGF-induced survival of chick sympathetic neurons by contact with a conditioned medium factor bound to the culture substrate.
    Brain Res. 1982 Sep;281(1):89-92 PMID: 7139342
  7. Induction of cell attachment and morphological differentiation in a pheochromocytoma cell line and embryonal sensory cells by the extracellular matrix.
    Dev Biol. 1982 Oct;93(2):285-300 PMID: 7141098
  8. Synthesis of type IV collagen and laminin in cultures of skeletal muscle cells and their assembly on the surface of myotubes.
    Dev Biol. 1982 Oct;93(2):344-54 PMID: 7141102
  9. Synthesis and localization of two sulphated glycoproteins associated with basement membranes and the extracellular matrix.
    J Cell Biol. 1982 Oct;95(1):197-204 PMID: 7142285
  10. Theory and methods for immunization in culture and monoclonal antibody production.
    J Immunol Methods. 1982 Sep 30;53(3):261-91 PMID: 6754815
  11. Isolation and characterization of type IV procollagen, laminin, and heparan sulfate proteoglycan from the EHS sarcoma.
    Biochemistry. 1982 Nov 23;21(24):6188-93 PMID: 6217835
  12. Sulfated proteoglycans synthesized by vascular endothelial cells in culture.
    J Biol Chem. 1983 Feb 10;258(3):2014-21 PMID: 6337150
  13. Nerve growth factor, laminin, and fibronectin promote neurite growth in human fetal sensory ganglia cultures.
    J Neurosci Res. 1982;8(2-3):179-93 PMID: 7154111
  14. Antiserum to a new neuronal growth factor: effects on neurite outgrowth.
    J Neurosci Res. 1982;8(2-3):289-302 PMID: 7154118
  15. The use of Tween 20 as a blocking agent in the immunological detection of proteins transferred to nitrocellulose membranes.
    J Immunol Methods. 1982 Dec 30;55(3):297-307 PMID: 6820029
  16. Production of laminin and fibronectin by Schwannoma cells: cell-protein interactions in vitro and protein localization in peripheral nerve in vivo.
    J Cell Biol. 1983 May;96(5):1218-26 PMID: 6341376
  17. In vivo and in vitro observations on laminin production by Schwann cells.
    Proc Natl Acad Sci U S A. 1983 Jun;80(12):3850-4 PMID: 6344090
  18. Neurite extension by peripheral and central nervous system neurons in response to substratum-bound fibronectin and laminin.
    Dev Biol. 1983 Jul;98(1):212-20 PMID: 6862106
  19. Synthesis of laminin and entactin by F9 cells induced with retinoic acid and dibutyryl cyclic AMP.
    J Biol Chem. 1983 Jun 25;258(12):7729-37 PMID: 6305950
  20. Characterization of protease-resistant fragments of laminin mediating attachment and spreading of rat hepatocytes.
    J Biol Chem. 1983 Jul 25;258(14):8922-7 PMID: 6408095
  21. Presence of nerve growth factor receptors and catecholamine uptake in subpopulations of chick sympathetic neurons: correlation with survival factor requirements in culture.
    Dev Biol. 1983 Sep;99(1):34-40 PMID: 6311648
  22. Spectrophotometric quantitation of silver grains eluted from autoradiograms.
    Anal Biochem. 1983 Sep;133(2):511-4 PMID: 6638510
  23. Laminin promotes neuritic regeneration from cultured peripheral and central neurons.
    J Cell Biol. 1983 Dec;97(6):1882-90 PMID: 6643580
  24. Glycosaminoglycans synthesized by cultured bovine corneal endothelial cells.
    J Cell Physiol. 1983 Dec;117(3):368-76 PMID: 6418749
  25. New neuronal growth factors.
    Annu Rev Neurosci. 1984;7:149-70 PMID: 6143526
  26. Distribution of entactin in the basement membrane of the rat mammary gland. Evidence for a non-epithelial origin.
    Exp Cell Res. 1984 May;152(1):240-54 PMID: 6538847
  27. Studies on extracellular matrix components that promote neurite outgrowth.
    Cold Spring Harb Symp Quant Biol. 1983;48 Pt 2:611-23 PMID: 6586379
  28. The production of a monoclonal antibody that blocks the action of a neurite outgrowth-promoting factor.
    Cold Spring Harb Symp Quant Biol. 1983;48 Pt 2:625-31 PMID: 6426848
  29. The heparin-binding domain of laminin is responsible for its effects on neurite outgrowth and neuronal survival.
    EMBO J. 1984 Jul;3(7):1463-8 PMID: 6745238
  30. Motoneurons purified by cell sorting respond to two distinct activities in myotube-conditioned medium.
    Dev Biol. 1984 Nov;106(1):194-210 PMID: 6489607
  31. The spatial control of ganglionic neurite growth by the substrate-associated material from conditioned medium: an experimental model of haptotaxis.
    J Neurosci. 1984 Nov;4(11):2823-9 PMID: 6502206
  32. Laminin is associated with the "neurite outgrowth-promoting factors" found in conditioned media.
    Proc Natl Acad Sci U S A. 1985 Apr;82(7):2183-7 PMID: 3856891
  33. Immunopurification and characterization of a neuronal heparan sulfate proteoglycan.
    J Neurosci. 1985 Jul;5(7):1842-50 PMID: 3160834
  34. PURIFICATION, COMPOSITION, AND MOLECULAR WEIGHT OF THE BETA-GALACTOSIDASE OF ESCHERICHIA COLI K12.
    J Biol Chem. 1965 Jun;240:2468-77 PMID: 14304855
  35. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels.
    Eur J Biochem. 1974 Jul 1;46(1):83-8 PMID: 4850204
  36. A simplified method for cyanogen bromide activation of agarose for affinity chromatography.
    Anal Biochem. 1974 Jul;60(1):149-52 PMID: 4850305
  37. Cell-to-substratum adhesion and guidance of axonal elongation.
    Dev Biol. 1975 May;44(1):92-101 PMID: 1132591
  38. The size and detergent binding of membrane proteins.
    J Biol Chem. 1975 Jul 25;250(14):5459-69 PMID: 1141239
  39. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography.
    Eur J Biochem. 1975 Aug 15;56(2):335-41 PMID: 1175627
  40. Stimulation of corneal endothelial cell proliferations in vitro by fibroblast and epidermal growth factors.
    Exp Eye Res. 1977 Jul;25(1):75-89 PMID: 302215
  41. Non-equivalence of conditioned medium and nerve growth factor for sympathetic, parasympathetic, and sensory neurons.
    Exp Cell Res. 1978 Apr;113(1):39-45 PMID: 25192
  42. Induction of neurite outgrowth by a conditioned-medium factor bound to the culture substratum.
    Proc Natl Acad Sci U S A. 1978 Oct;75(10):5210-3 PMID: 217018
  43. The coating of bovine and rabbit corneas denuded of their endothelium with bovine corneal endothelial cells.
    Exp Eye Res. 1979 Mar;28(3):249-65 PMID: 374105
  44. Laminin--a glycoprotein from basement membranes.
    J Biol Chem. 1979 Oct 10;254(19):9933-7 PMID: 114518
  45. Basement membrane glycoprotein laminin binds to heparin.
    FEBS Lett. 1980 Jul 28;116(2):243-6 PMID: 6250887
  46. Neurite outgrowth induced by the substrate associated material from nonneuronal cells.
    Dev Biol. 1980 Sep;79(1):247-52 PMID: 7409323
  47. Characterization of pepsin fragments of laminin in a tumor basement membrane. Evidence for the existence of related proteins.
    Hoppe Seylers Z Physiol Chem. 1980 Nov;361(11):1651-60 PMID: 6778807
  48. Polyornithine-attached neurite-promoting factors (PNPFs). Culture sources and responsive neurons.
    Brain Res. 1981 Feb 9;206(1):129-44 PMID: 7470880
  49. Regulation of the elongating nerve fiber.
    Curr Top Dev Biol. 1980;16:165-206 PMID: 6162612
  50. Characterization of a neuronal growth factor from mouse heart-cell-conditioned medium.
    Dev Biol. 1981 Feb;82(1):56-68 PMID: 7227638
  51. Entactin, a novel basal lamina-associated sulfated glycoprotein.
    J Biol Chem. 1981 May 25;256(10):5209-14 PMID: 6262321
  52. The production and localization of laminin in cultured vascular and corneal endothelial cells.
    J Cell Physiol. 1981 May;107(2):171-83 PMID: 7251679
  53. Neurite-promoting factor in conditioned medium from RN22 Schwannoma cultures: bioassay, fractionation, and properties.
    J Neurochem. 1981 Sep;37(3):759-67 PMID: 7276956
  54. Promotion of retinal neurite outgrowth by substratum-bound fibronectin.
    Dev Biol. 1981 Aug;86(1):179-88 PMID: 7286392
  55. Shapes, domain organizations and flexibility of laminin and fibronectin, two multifunctional proteins of the extracellular matrix.
    J Mol Biol. 1981 Jul 25;150(1):97-120 PMID: 6795355
  56. Protease resistance and conformation of laminin.
    Eur J Biochem. 1982 Mar;123(1):63-72 PMID: 7040076
  57. Binding of the basement-membrane glycoprotein laminin to glycosaminoglycans. An affinity-chromatography study.
    Biochem J. 1981 Dec 1;199(3):699-704 PMID: 7340825
  58. Structure of the pericellular matrix: association of heparan and chondroitin sulfates with fibronectin-procollagen fibers.
    Cell. 1982 Mar;28(3):663-71 PMID: 6210440
  59. Studies on the biosynthesis of laminin by murine parietal endoderm cells.
    Eur J Biochem. 1981 Sep;119(1):189-97 PMID: 7341241
  60. Fibronectin: purification, immunochemical properties, and biological activities.
    Methods Enzymol. 1982;82 Pt A:803-31 PMID: 7078457
  61. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.101.3.898
Published
1985-09-00
Pages
898-913
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2113739
Subset
IM
Grants
NIGMS NIH HHS · GM07618 · United States
NINDS NIH HHS · NS19090 · United States
External Links
PubMed source DOI: 10.1083/jcb.101.3.898 PMC Full Text View journal details
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