LOW NIDOGEN AFFINITY OF LAMININ-5 CAN BE ATTRIBUTED TO 2 SERINE RESIDUES IN EGF-LIKE MOTIF GAMMA-2III4

High affinity nidogen binding of laminin-1 (chain composition alpha 1 beta 1 gamma 1) has been previously mapped to a single EGF-like motif gamma 1III4 of its gamma 1 chain, Two more isoforms, laminin-5 (alpha 3 beta 3 gamma 2) and laminin-7 (alpha 3 beta 2 gamma 1), show low and high binding activity, respectively, indicating that the gamma 2 chai n is of low affinity, This was confirmed by recombinant production of the homologous EGF-like motif gamma 2III4 of the gamma 2 chain, which has a 100,000-fold lower binding activity than gamma 1III4, The crucia l heptapeptide binding sequence Asn-lle-Asp-Pro-Asn-Ala-Val of gamma 1 III4 is modified in gamma 2III4 by replacing both the central Asn and Val by Ser, Changing these replacements to Asn and Val by site-directe d mutagenesis enhanced the activity of gamma 2III4 to a level which wa s only 5-fold lower than that of gamma 1III4. Despite their high seque nce identity (77%) motifs gamma 1III4 and gamma 2III4 were also shown to differ considerably in immunological epitopes, This indicates disti nctly different functions for laminins which differ in the gamma chain isoform.