Crystal structure of a phage library-derived single-chain Fv fragment complexed with turkey egg-white lysozyme at 2.0 A resolution.
The three-dimensional structure of the single-chain Fv fragment 1F9 in complex with turkey egg-white lysozyme (TEL) has been determined to a nominal resolution of 2.0 A by X-ray diffraction. The scFv fragment 1F9 was derived from phage-display libraries in two steps and binds both hen and turkey egg-white lysozyme, although the level of binding affinity is two orders of magnitude greater for the turkey lysozyme. The comparison of the crystal structure with a model of the single-chain Fv fragment 1F9 in complex with hen egg-white lysozyme (HEL) reveals that in the latter a clash between Asp101 in lysozyme and Trp98 of the complementarity determining region H3 of the heavy chain variable domain occurs. This is the only explanation apparent from the crystal structure for the better binding of TEL compared to HEL. The binding site topology on the paratope is not simply a planar surface as is usually found in antibody-protein interfaces, but includes a cleft between the light chain variable domain and heavy chain variable domain large enough to accommodate a loop from the lysozyme. The scFv fragment 1F9 recognizes an epitope on TEL that differs from the three antigenic determinants recognized in other known crystal structures of monoclonal antibodies in complex with lysozyme.
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Abteilung Proteinstrukturforschung, Institut für Biochemie des Universitätsklinikums Charité der Humboldt-Universität zu Berlin, Monbijoustr. 2, Berlin, 10117, Germany.
Keitel
T
T
Küttner
G
G
Wessner
H
H
Scholz
C
C
Hahn
M
M
Höhne
W
W
D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D000937:Antigen-Antibody Reactions; D001435:Bacteriophages / Q000235:genetics; D018360:Crystallography, X-Ray; D004527:Egg Proteins / Q000737:chemistry*; D018604:Epitope Mapping; D007135:Immunoglobulin Variable Region / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009113:Muramidase / Q000737:chemistry*; D014422:Turkeys
D000596:Amino Acids; D004527:Egg Proteins; D007135:Immunoglobulin Variable Region; D009113:Muramidase