Laminin-121--recombinant expression and interactions with integrins.

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.

• Authors: Takako Sasaki, Junichi Takagi, Camilla Giudici, Yoshihiko Yamada, Eri Arikawa-Hirasawa, Rainer Deutzmann, Rupert Timpl, Arnoud Sonnenberg, Hans Peter Bächinger, David Tonge
• Journal: Matrix biology : journal of the International Society for Matrix Biology (Matrix Biol) Vol. 29 Issue 6 Pg. 484-93 (Jul 2010) ISSN: 1569-1802 [Electronic] Netherlands
• PMID: 20566382 (Publication Type: Comparative Study, Journal Article, Research Support, N.I.H., Intramural, Research Support, Non-U.S. Gov't)
• Copyright: Published by Elsevier B.V.

Chemical References

• Integrins
• Laminin
• Recombinant Proteins
• heparin-sepharose
• laminin beta2
• laminin A
• Sepharose
• integrin alpha7beta1

Topics

• Animals
• Cell Line
• Chromatography, Gel
• Circular Dichroism
• HEK293 Cells
• Humans
• In Vitro Techniques
• Integrins (genetics, metabolism)
• Kidney (cytology)
• Laminin (chemistry, genetics, isolation & purification, metabolism)
• Mice
• Recombinant Proteins (metabolism, ultrastructure)
• Sepharose (analogs & derivatives, chemistry)
• Transfection