Structural changes in the NH2-terminal domain of fibronectin upon interaction with heparin. Relationship to matrix-driven translocation.
The effects of heparin and various related polysaccharides on the circular dichroic spectra of fibronectin and its 31-kDa NH2-terminal tryptic fragment were studied. These effects were evaluated with respect to (i) spectral features of the native proteins that are sensitive to pH denaturation and breaking of disulfide bonds, (ii) sensitivity of spectral changes to Ca2+, and (iii) the fibronectin-dependent interfacial interaction known as "matrix-driven translocation." We found that native heparin causes an attenuation of the positive CD peak at 228 nm with both the intact protein and the fragment, and causes a small but reproducible red shift in the spectrum of the fragment. All of these changes are analogous to spectral changes seen with denaturation or reduction of the proteins. In contrast to the situation with the intact protein, the heparin-induced spectral changes in the fragment were abolished in the presence of 10 mM Ca2+. Desulfation of heparin lessened or destroyed its ability to induce these changes, and carboxymethylated heparin and dextran sulfate induced different kinds of spectral alterations. Fibronectin and heparin determinants required for the induction of the characteristic spectral shift of the NH2-terminal domain corresponded to those required for matrix-driven translocation, suggesting that the associated conformational change in fibronectin plays a role in this biophysical effect.
Khan
M Y
MY
Department of Anatomy, New York Medical College, Valhalla 10595.
Jaikaria
N S
NS
Frenz
D A
DA
Villanueva
G
G
Newman
S A
SA
D000345:Affinity Labels / Q000378:metabolism; D000465:Algorithms; D000595:Amino Acid Sequence; D002942:Circular Dichroism; D016264:Dextran Sulfate; D003911:Dextrans / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism*; D008970:Molecular Weight; D011487:Protein Conformation
D000345:Affinity Labels; D003911:Dextrans; D005353:Fibronectins; D006493:Heparin; D016264:Dextran Sulfate