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PMID:20566382

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Citation

Sasaki, T, Takagi, J, Giudici, C, Yamada, Y, Arikawa-Hirasawa, E, Deutzmann, R, Timpl, R, Sonnenberg, A, Bächinger, HP and Tonge, D (2010) Laminin-121--recombinant expression and interactions with integrins. Matrix Biol. 29:484-93

Abstract

Laminin-121, previously referred as to laminin-3, was expressed recombinantly in human embryonic kidney (HEK) 293 cells by triple transfection of full-length cDNAs encoding mouse laminin α1, β2 and γ1 chains. The recombinant laminin-121 was purified using Heparin-Sepharose followed by molecular sieve chromatography and shown to be correctly folded by electron microscopy and circular dichroism (CD). The CD spectra of recombinant laminin-121 were very similar to those of laminin-111 isolated from Engelbreth-Holm-Swarm tumor (EHS-laminin) but its T(m) value was smaller than EHS-laminin and recombinant lamnin-111 suggesting that the replacement of the β chain reduced the stability of the coiled-coil structure of laminin-121. Its binding to integrins was compared with EHS-laminin, laminin-3A32 purified from murine epidermal cell line and recombinantly expressed laminins-111, -211 and -221. Laminin-121 showed the highest affinity to α6β1 and α7β1 integrins and furthermore, laminin-121 most effectively supported neurite outgrowth. Together, this suggests that the β2 laminins have higher affinity for integrins than the β1 laminins.

Links

PubMed PMC2939186 Online version:10.1016/j.matbio.2010.05.004

Keywords

Animals; Cell Line; Chromatography, Gel; Circular Dichroism; HEK293 Cells; Humans; Integrins/genetics; Integrins/metabolism; Kidney/cytology; Laminin/chemistry; Laminin/genetics; Laminin/isolation & purification; Laminin/metabolism; Mice; Recombinant Proteins/metabolism; Recombinant Proteins/ultrastructure; Sepharose/analogs & derivatives; Sepharose/chemistry; Transfection

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

MOUSE:LAMB2

enables

GO:0005178: integrin binding

ECO:0000353: physical interaction evidence used in manual assertion

PR:P09055
PR:Q61738
PR:Q61739

F

Seeded From UniProt

complete

MOUSE:LAMB2

acts_upstream_of_or_within

GO:0031175: neuron projection development

ECO:0000314: direct assay evidence used in manual assertion

P

  • occurs_in:(CL:0000540)
  • occurs_in:(EMAPA:16668)

Seeded From UniProt

complete

MOUSE:LAMB2

part_of

GO:0005608: laminin-3 complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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