Conformational flexibility and crystallization of tandemly linked type III modules of human fibronectin.
Fibronectin is a large cell adhesion molecule that is composed of several functional domains. The cell-binding domain that binds to cell surface integrins consists of repeated homologous type III modules. In this study, recombinant fragments from the cell-binding domain of human fibronectin that participate in a newly characterized fibronectin-fibronectin interaction with FNIII1 were crystallized. In each case, the crystals had more than one fibronectin fragment in the asymmetric unit. Crystals of FNIII10-11 grew in the space group C2 with a = 117.1 A, b = 38.6 A, c = 80.6 A, beta = 97.2 degrees, and two molecules in the asymmetric unit. These crystals diffracted to 2.5 A resolution. Fragment FNIII8-11 and a shorter fragment, FNIII8-10, crystallized in hexagonal space groups with large unit cells and two to four molecules per asymmetric unit. Even very large crystals of these fragments did not diffract beyond 4 A. The crystal packing for this collection of fibronectin fragments suggests conformational flexibility between linked type III modules. The functional relevance of this flexibility for elongated versus compact models of the cell-binding domain of fibronectin is discussed.
Lombardo
A
A
La Jolla Cancer Center, Burnham Institute, California 92037, USA.
Wang
Y
Y
Ni
C Z
CZ
Dai
X
X
Dickinson
C D
CD
Kodandapani
R
R
Chiang
S
S
White
C A
CA
Pio
F
F
Xuong
N H
NH
Hamlin
R C
RC
Ruoslahti
E
E
Ely
K R
KR
D000645:Ammonium Sulfate; D001665:Binding Sites; D011232:Chemical Precipitation; D003460:Crystallization; D005353:Fibronectins / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D016023:Integrins / Q000378:metabolism; D008278:Magnesium Sulfate; D010446:Peptide Fragments / Q000737:chemistry; D011092:Polyethylene Glycols; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D014961:X-Ray Diffraction
D005353:Fibronectins; D016023:Integrins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D008278:Magnesium Sulfate; D000645:Ammonium Sulfate