Targeting of the immunoglobulin-binding domain of protein A to the extracellular matrix using a minifibronectin expression vector.
A truncated form of fibronectin consisting of the N-terminal 70-kDa and C-terminal 37-kDa regions, referred to as "minifibronectin," retains the ability to assemble into the extracellular matrix, even though it lacks the central approximately 120-kDa region containing most of the type III modules (Ichihara-Tanaka, K., Titani, K., and Sekiguchi, K., FEBS Lett. 299, 155-158, 1992). Taking advantage of the matrix assembly activity of minifibronectin, we developed a novel method to target non-matrix proteins to the extracellular matrix by inserting them between the N-terminal 70-kDa and the C-terminal 37-kDa regions of minifibronectin. Using the immunoglobulin-binding domain of Staphylococcal protein A as a model, we demonstrated that the bacterial protein expressed in mouse L cells as a chimeric protein with minifibronectin is secreted as disulfide-bonded dimers and successfully deposited onto the extracellular matrix of transfected cells. The chimeric protein retained the immunoglobulin-binding activity not only in solution but also after deposition at the matrix. This targeting strategy we developed will provide a means to manipulate the biological functions of the extracellular matrix through targeting of a wide variety of non-matrix proteins.
Matsuyama
S
S
Research Institute, Osaka Medical Center for Maternal and Child Health.
Ichihara-Tanaka
K
K
Sekiguchi
K
K
D000818:Animals; D001665:Binding Sites; D002448:Cell Adhesion / Q000502:physiology; D004220:Disulfides / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism / Q000502:physiology*; D005353:Fibronectins / Q000378:metabolism*; D005822:Genetic Vectors; D006041:Goats; D006801:Humans; D007136:Immunoglobulins / Q000378:metabolism* / Q000648:ultrastructure; D007739:L Cells; D051379:Mice; D009842:Oligopeptides / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013205:Staphylococcal Protein A / Q000378:metabolism* / Q000648:ultrastructure; D014162:Transfection
D004220:Disulfides; D005353:Fibronectins; D007136:Immunoglobulins; D009842:Oligopeptides; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; C047981:arginyl-glycyl-aspartic acid