Laminin isoforms containing the gamma3 chain are unable to bind to integrins due to the absence of the glutamic acid residue conserved in the C-terminal regions of the gamma1 and gamma2 chains.
Laminins are the major cell adhesive proteins in basement membranes, and consist of three subunits termed alpha, beta, and gamma. Recently, we found that the Glu residue at the third position from the C termini of the gamma1 and gamma2 chains is critically involved in integrin binding by laminins. However, the gamma3 chain lacks this Glu residue, suggesting that laminin isoforms containing the gamma3 chain may be unable to bind to integrins. To address this possibility, we expressed the E8 fragment of laminin-213 and found that it was incapable of binding to integrins. Similarly, the E8 fragment of laminin-113 was expressed and also found to be inactive in binding to integrins, confirming the distinction between the integrin binding activities of gamma3 chain-containing isoforms and those containing the gamma1 or gamma2 chain. To further address the importance of the Glu residue, we swapped the C-terminal four amino acids of the gamma3 chain with the C-terminal nine amino acids of the gamma1 chain, which contain the Glu residue. The resulting chimeric E8 fragment of laminin-213 became fully active in integrin binding, whereas replacement with the nine amino acids of the gamma1 chain after substitution of Gln for the conserved Glu residue failed to restore the integrin binding activity. These results provide both loss-of-function and gain-of-function evidence that laminin isoforms containing the gamma3 chain are unable to bind to integrins due to the absence of the conserved Glu residue, which should play a critical role in integrin binding by laminins.
Ido
Hiroyuki
H
Laboratory of Extracellular Matrix Biochemistry, Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
Ito
Shunsuke
S
Taniguchi
Yukimasa
Y
Hayashi
Maria
M
Sato-Nishiuchi
Ryoko
R
Sanzen
Noriko
N
Hayashi
Yoshitaka
Y
Futaki
Sugiko
S
Sekiguchi
Kiyotoshi
K
D000595:Amino Acid Sequence; D002448:Cell Adhesion; D004305:Dose-Response Relationship, Drug; D018698:Glutamic Acid / Q000737:chemistry*; D006801:Humans; D020014:K562 Cells; D007797:Laminin / Q000737:chemistry*; D008954:Models, Biological; D008969:Molecular Sequence Data; D011485:Protein Binding; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary
C400567:LAMC2 protein, human; C504218:LAMC3 protein, human; D007797:Laminin; D020033:Protein Isoforms; C089141:laminin gamma 1; D018698:Glutamic Acid