Characterization of commercial laminin preparations from human placenta in comparison to recombinant laminins 2 (alpha2beta1gamma1), 8 (alpha4beta1gamma1), 10 (alpha5beta1gamma1).
Laminins, a family of large heterotrimeric (alphabetagamma) proteins, are major components of basement membranes implicated in a variety of cellular functions. Different commercial laminin preparations isolated from human placenta have been widely used in functional studies but their molecular properties are poorly known. In the present study, we characterized several of these preparations by ELISA, silver staining and Western blotting, in comparison to mouse laminin 1 (alpha1beta1gamma1), and recombinant human laminins 2 (alpha2beta1gamma1), 8 (alpha4beta1gamma1) and 10 (alpha5beta1gamma1). The cell migration-promoting activity of different batches was also tested. The placenta laminin preparations differed from one another and consisted of highly fragmented proteins, a mixture of laminin isoforms, and/or contaminating fibronectin. Major functional differences between batches were also observed, reflecting molecular heterogeneity. Previous data obtained in functional studies using these preparations need to be interpreted with caution and may require revision, and future functional studies demand prior molecular characterization of the laminins, particularly their alpha-chain.
Wondimu
Zenebech
Z
Department of Odontology, Karolinska Institutet at Huddinge, SE-141 04 Stockholm, Sweden.
Gorfu
Gezahegn
G
Kawataki
Tomoyuki
T
Smirnov
Sergei
S
Yurchenco
Peter
P
Tryggvason
Karl
K
Patarroyo
Manuel
M
D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007797:Laminin / Q000235:genetics* / Q000302:isolation & purification; D051379:Mice; D010920:Placenta / Q000502:physiology*; D011247:Pregnancy; D011994:Recombinant Proteins / Q000737:chemistry*
D007797:Laminin; D011994:Recombinant Proteins; C119013:laminin 10; C110308:laminin 8; C102645:laminin alpha 2