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

https://www.ncbi.nlm.nih.gov/pubmed/28745050

dc.identifier.eissn

1520-4995

dc.identifier.uri

https://hdl.handle.net/10161/15431

dc.language

eng

dc.publisher

American Chemical Society (ACS)

dc.relation.ispartof

Biochemistry

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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

https://www.ncbi.nlm.nih.gov/pubmed/28745050

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

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Institutes and Centers

pubs.organisational-group

School of Medicine

pubs.publication-status

Published online

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