Purification, crystallization and preliminary crystallographic studies of a two fibronectin type-III domain segment from chicken tenascin encompassing the heparin- and contactin-binding regions.
A fragment of chicken tenascin consisting of fibronectin type-III domains 5 and 6 has been expressed in Escherichia coli. After modifying a previously reported purification protocol, an electrophoretically homogeneous recombinant protein was obtained from which various crystal forms could be grown under identical conditions. Only one form was suitable for structure determination. These crystals belong to space group P21, with unit-cell parameters a = 45.2, b = 57.9, c = 72.2 A, beta = 91.4 degrees, and diffract to at least 2.6 A resolution using synchrotron radiation. From density measurements of the crystals, it was found that there are two molecules in the asymmetric unit. Diffraction data of native, two platinum-derivative and one palladium-derivative crystals were collected.
Bisig
D
D
Laboratorium für Biochemie I, Swiss Federal Institute of Technology (ETH), CH-8092 Zürich, Switzerland.
Weber
P
P
Vaughan
L
L
Winterhalter
K H
KH
Piontek
K
K
D000818:Animals; D001665:Binding Sites; D015816:Cell Adhesion Molecules, Neuronal / Q000737:chemistry* / Q000378:metabolism*; D002645:Chickens; D058966:Contactins; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006493:Heparin / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D046911:Macromolecular Substances; D010446:Peptide Fragments / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D019063:Tenascin / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism
D015816:Cell Adhesion Molecules, Neuronal; D058966:Contactins; D046911:Macromolecular Substances; D010446:Peptide Fragments; D011994:Recombinant Proteins; D019063:Tenascin; D006493:Heparin