Abstract
The structure of human plasma fibronectin in 50 mM Tris-HCl buffer, pH 7.4, containing varying concentrations of NaCl, has been investigated using the small-angle X-ray method.
Below 0.3 M NaCl the overall structure of the molecule is disc-shaped; at 0 M NaCl the axial ratio of the disc is about 1:7 and between 0.1 M to 0.3 M it is slightly more asymmetric, with an axial ratio of 1:10.
At about 0.3 M NaCl there is a reversible transition to a more open structure, and, from 0.3 M up to 1.1 M NaCl the small-angle X-ray data can be explained by models consisting of ensembles of flexible, non-overlapping, bead-chains generated by a Monte Carlo procedure. Within this concentration range there is a gradual increase in the stiffness of the chains, as well as a decrease in bead radius, which indicates that the molecule becomes more open when the NaCl concentration is increased.
The transition to a more open structure is also demonstrated by the average radius of gyration which increases gradually from 8.26 nm at 0 M NaCl to 8.75 nm at physiological or near-physiological conditions, and up to 16.2 nm at 1.1 M NaCl.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- hpFN:
-
human plasma fibronectin
- SAXS:
-
smallangle X-ray scattering
- Tris :
-
tris (hydroxymethyl) aminomethane
- DTT:
-
dithiothreitol
- BA:
-
benzamidine hydrochloride
- PMSF:
-
phenylmethylsulfonyl fluoride
References
Akiyama SK, Yamada KM (1986) Fibronectin. Adv Enzymol 59:1–57
Creighton TE (1984) Proteins, structures and molecular properties. WH Freeman, New York
Debye P (1915) Zerstreuung von Röntgenstrahlen. Ann Physik 46:809–823
Debye P (1947) Molecular-weight determination by light scattering. J Phys Colloid Chem 51:18–32
Engel J, Odermatt E, Engel A, Madri JA, Furthmayr H, Rohde H, Timpl R (1981) Shapes, domain organizations and flexibility of laminin and fibronectin, two multifunctional proteins of the extracellular matrix. J Mol Biol 150:97–120
Erickson HP, Carrell NA (1983) Fibronectin in extended and compact conformations. Electron microscopy and sedimentation analysis. J Biol Chem 258:14539–14544
Goldstein H (1959) Classical mechanics. Addison-Wesley, Reading, Massachusetts, USA
Guinier A, Fournet G (1955) Small-angle scattering of X-rays. John Wiley, New York
Kornblihtt AR, Umezawa K, Vibe-Pedersen K, Baralle FE (1985) Primary structure of human fibronectin: differential splicing may generate at least 10 polypeptides from a single gene. EMBO J 4:1755–1759
Koteliansky VE, Glukhova MA, Bejanian MV, Smirnov VN, Filimonov VV, Zalite OM, Venyaminov SY (1981) A study of the structure of fibronectin. Eur J Biochem 119:619–624
Kratky O, Pilz I, Schmitz PJ (1966) Absolute intensity measurement of small angle X-ray scattering by means of a standard sample. J Colloid Interface Sci 21:24–34
Kratky O, Leopold H, Stabinger H (1969) Dichtemessungen an Flüssigkeiten und Gasen auf 10-6 g/cm3 bei 0.6 cm3 Präparatvolumen. Z Angew Physik 27:273–277
Kratky O, Leopold H, Seidler H-P (1971) Röntgen-Kleinwinkelkamera mit Monitor. Z Angew Physik 31:49–50
Lai C-S, Tooney NM, Ankel EG (1984) Structure and flexibility of plasma fibronectin in solution: Electron spin resonance spin-label, circular dichroism, and sedimentation studies. Biochemistry 23:6393–6397
Markovic Z, Lustig A, Engel J, Richter H, Hörmann H (1983) Shape and stability of fibronectin in solutions of different pH and ionic strength. Hoppe-Seyler's Z Physiol Chem 364: 1795–1804
McDonagh J, Ed (1985) Plasma fibronectin, structure and function. Marcel Dekker, New York.
Mosesson MW, Chen AB, Huseby RM (1975) The coldinsoluble globulin of human plasma: Studies of its essential structural features. Biochim Biophys Acta 386:509–524
NAG-Fortran manual (1983) Numerical Algorithms Group Ltd, NAG Central office, Mayfield House, 256 Banbury Road, Oxford OX2 7DE, UK
von Neumann J (1951) Various techniques used in connection with random digits. Natl Bur Stand, Appl Math Series 12:36–38
Odermatt E, Engel J, Richter H, Hörmann H (1982) Shape, conformation and stability of fibronectin fragments determined by electron microscopy, circular dichroism and ultracentrifugation. J Mol Biol 159:109–123
Price TM, Rudee ML, Pierschbacher M, Ruoslahti E (1982) Structure of fibronectin and its fragments in electron microscopy. Eur J Biochem 129:359–363
Rocco M, Carson M, Hantgan R, McDonagh J, Hermans J (1983) Dependence of the shape of the plasma fibronectin molecule on solvent composition. Ionic strength and glycerol content. J Biol Chem 258:14545–14549
Rocco M, Infusini E, Daga MG, Gogioso L, Cuniberti C (1987) Models of fibronectin. EMBO J 6:2343–2349
Schellman JA (1974) Flexibility of DNA. Biopolymers 13:217–226
Sjöberg B (1978) A general, weighted least-squares method for the evaluation of small-angle X-ray data without desmearing. J Appl Crystallogr 11:73–79
Sjöberg B, Pap S, Österlund E, Österlund K, Vuento M, Kjems J (1987) Solution structure of human plasma fibronectin using small-angle X-ray and neutron scattering at physiological pH and ionic strength. Arch Biochem Biophys 255:347–353
Skorstengaard K, Jensen MS, Sahl P, Petersen TE, Magnusson S (1986) Complete primary structure of bovine plasma fibronectin. Eur J Biochem 161:441–453
Vuento M, Vaheri A (1979) Purification of fibronectin from human plasma by affinity chromatography under nondenaturing conditions. Biochem J 183:331–337
Yamada KM (1983) Cell surface interactions with extracellular materials. Annu Rev Biochem 52:761–799
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Sjöberg, B., Eriksson, M., Österlund, E. et al. Solution structure of human plasma fibronectin as a function of NaCl concentration determined by small-angle X-ray scattering. Eur Biophys J 17, 5–11 (1989). https://doi.org/10.1007/BF00257140
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF00257140