Fibronectin Conformation and Assembly: Analysis of Fibronectin Deletion Mutants and Fibronectin Glomerulopathy (GFND) Mutants.
dc.contributor.author | Ohashi, Tomoo | |
dc.contributor.author | Lemmon, Christopher A | |
dc.contributor.author | Erickson, Harold P | |
dc.coverage.spatial | United States | |
dc.date.accessioned | 2017-09-01T21:31:55Z | |
dc.date.available | 2017-09-01T21:31:55Z | |
dc.date.issued | 2017-08-11 | |
dc.description.abstract | To study fibronectin (FN) conformation and assembly, we generated several deletion mutants: FNΔ(I)1-5, FNΔ(III)1-3, FNΔ(III)4-8, and FNΔ(III)11-14. A monomeric form, FNmono, which lacked the C-terminal dimerization region, was also created. FNtnA-D was generated by swapping FNIII domains 1-8 in FNΔ(III)11-14 with seven FNIII domains from tenascin-C. The conformations of these mutants were analyzed by glycerol gradient sedimentation under low-salt (20 mM NaCl) and high-salt (200 mM NaCl) conditions. Surprisingly, most of the mutants showed a compact conformation under low-salt conditions, except for FNtnA-D. When we tested these mutants in cell culture, FNΔ(I)1-5, FNΔ(III)1-3, and FNtnA-D were unable to form a matrix. Interestingly, FNΔ(III)1-3 and FNtnA-D were capable of co-assembly with full-length FN, while FNΔ(I)1-5 was not. This indicates that the segment (I)1-5 is crucial for matrix assembly and segment (III)1-3 is also important. Mutations in FN are associated with glomerulopathy, but when we studied mutant proteins, the single-nucleotide mutations had only minor effects on conformation and matrix assembly. The mutations may destabilize their FNIII domains or generate dimers of dimers by disulfide cross-linking. | |
dc.identifier | ||
dc.identifier.eissn | 1520-4995 | |
dc.identifier.uri | ||
dc.language | eng | |
dc.publisher | American Chemical Society (ACS) | |
dc.relation.ispartof | Biochemistry | |
dc.relation.isversionof | 10.1021/acs.biochem.7b00589 | |
dc.title | Fibronectin Conformation and Assembly: Analysis of Fibronectin Deletion Mutants and Fibronectin Glomerulopathy (GFND) Mutants. | |
dc.type | Journal article | |
duke.contributor.orcid | Erickson, Harold P|0000-0002-9104-8987 | |
pubs.author-url | ||
pubs.organisational-group | Basic Science Departments | |
pubs.organisational-group | Biochemistry | |
pubs.organisational-group | Cell Biology | |
pubs.organisational-group | Duke | |
pubs.organisational-group | Duke Cancer Institute | |
pubs.organisational-group | Institutes and Centers | |
pubs.organisational-group | School of Medicine | |
pubs.publication-status | Published online |