| First Author | Sasaki T | Year | 1995 |
| Journal | J Mol Biol | Volume | 245 |
| Issue | 3 | Pages | 241-50 |
| PubMed ID | 7844816 | Mgi Jnum | J:22417 |
| Mgi Id | MGI:70289 | Doi | 10.1006/jmbi.1994.0020 |
| Citation | Sasaki T, et al. (1995) Structural characterization of two variants of fibulin-1 that differ in nidogen affinity. J Mol Biol 245(3):241-50 |
| abstractText | Two C-terminal variants C and D of mouse fibulin-1 were purified from the culture medium of stably transfected human kidney cell clones. They showed, after rotary shadowing, a dumbbell-like structure of about 33 nm in length. Pepsin digestion demonstrated stability of the disulfide-bonded domains 1 (anaphylatoxin-like) and II (multiple EGF-like motifs) but not for domain III which is different in the variants. A close similarity of the variants was observed in immunochemical assays indicating that domain III epitopes are not very antigenic. Binding analysis in solid phase assays demonstrated for variant C a 100-fold stronger binding to the basement membrane protein nidogen than for variant D. Both interactions were sensitive to EDTA. Surface plasmon resonance assays confirmed this difference and showed KD = 60 nM for variant C and KD > 1 microM for variant D. Lower binding activities and smaller differences between both variants were observed for the calcium-dependent binding to fibronectin, laminin-1 and collagen IV. Self aggregation into nest-like oligomers was observed at high concentrations of fibulin-1 which was not sensitive to EDTA. |
