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Inhibition of glycosaminoglycan modification of perlecan domain I by site‐directed mutagenesis changes protease sensitivity and laminin‐1 binding activity
Author(s) -
Sasaki Takako,
Costell Mercedes,
Mann Karlheinz,
Timpl Rupert
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)01063-1
Subject(s) - proteases , mutagenesis , perlecan , site directed mutagenesis , glycosaminoglycan , chemistry , biochemistry , protease , laminin , recombinant dna , heparan sulfate , binding site , serine protease , mutation , enzyme , mutant , gene , extracellular matrix
Glycosaminoglycan attachment to perlecan domain I (173 residues) was completely prevented by site‐directed mutagenesis of Ser‐65, Ser‐71 and Ser‐76 as shown by recombinant production in mammalian cells. This did not interfere with the proper folding of the domain's SEA module but enhanced its sensitivity to neutral proteases. Lack of substitution also abolished binding to the two major heparin binding sites of laminin‐1.