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Comparison of the crystallization and crystal packing of two Fab single-site mutant protein L complexes

Comparison of the crystallization and crystal packing of two Fab single-site mutant protein L complexes
Comparison of the crystallization and crystal packing of two Fab single-site mutant protein L complexes
Protein L from Peptostreptococcus magnus (PpL) is a multidomain protein composed of four or five immunoglobulin-binding domains that target the light chain of a large repertoire of human and murine antibodies. Thus, a single domain of this protein can be used to aid the crystallization of Fab, free or complexed to their antigen when it is not possible to obtain crystals without it. Each wild-type PpL domain has two light-chain binding sites that target the same region of the light chain and can thus bring together two Fab-antigen complexes within the crystal lattice. In this context the small PpL domain is sandwiched between two Fab and cannot participate in crystal contacts, thus mutants are unlikely to increase the chances of crystallizing a particular complex. However, it is possible to design mutants that can bind at only one site by making use of the crystal structures obtained so far. Such mutants will have a free surface that can participate in crystal contacts and that can be modified to improve its crystal contact-forming properties. Here, a comparison of two single-site mutants that differ at three different positions is reported. In both mutants two different tryptophan residues participate in crystal-packing interactions, suggesting that this residue may be particularly interesting for enhancing crystal-contact formation.
0907-4449
750-754
Granata, V.
d8ab56a3-6226-49eb-bb0e-c82bc6d82a9f
Housden, N.G.
5fc2b5f1-a7da-4f20-b794-d7d8ef597d5d
Harrison, S.
0e3bb8ab-7b95-4686-9b74-f889065ad068
Jolivet-Reynaud, C.
84f997e7-3528-446d-b367-168a5e799afa
Gore, M.G.
7bd6db4b-c5a2-4206-8666-b92208ba7979
Stura, E.A.
43c87eeb-ec69-41bc-8a00-bf8f6e354bb4
Granata, V.
d8ab56a3-6226-49eb-bb0e-c82bc6d82a9f
Housden, N.G.
5fc2b5f1-a7da-4f20-b794-d7d8ef597d5d
Harrison, S.
0e3bb8ab-7b95-4686-9b74-f889065ad068
Jolivet-Reynaud, C.
84f997e7-3528-446d-b367-168a5e799afa
Gore, M.G.
7bd6db4b-c5a2-4206-8666-b92208ba7979
Stura, E.A.
43c87eeb-ec69-41bc-8a00-bf8f6e354bb4

Granata, V., Housden, N.G., Harrison, S., Jolivet-Reynaud, C., Gore, M.G. and Stura, E.A. (2005) Comparison of the crystallization and crystal packing of two Fab single-site mutant protein L complexes. Acta Crystallographica Section D: Biological Crystallography, 61, 750-754. (doi:10.1107/S0907444905007110).

Record type: Article

Abstract

Protein L from Peptostreptococcus magnus (PpL) is a multidomain protein composed of four or five immunoglobulin-binding domains that target the light chain of a large repertoire of human and murine antibodies. Thus, a single domain of this protein can be used to aid the crystallization of Fab, free or complexed to their antigen when it is not possible to obtain crystals without it. Each wild-type PpL domain has two light-chain binding sites that target the same region of the light chain and can thus bring together two Fab-antigen complexes within the crystal lattice. In this context the small PpL domain is sandwiched between two Fab and cannot participate in crystal contacts, thus mutants are unlikely to increase the chances of crystallizing a particular complex. However, it is possible to design mutants that can bind at only one site by making use of the crystal structures obtained so far. Such mutants will have a free surface that can participate in crystal contacts and that can be modified to improve its crystal contact-forming properties. Here, a comparison of two single-site mutants that differ at three different positions is reported. In both mutants two different tryptophan residues participate in crystal-packing interactions, suggesting that this residue may be particularly interesting for enhancing crystal-contact formation.

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Published date: 1 June 2005

Identifiers

Local EPrints ID: 56424
URI: https://eprints.soton.ac.uk/id/eprint/56424
ISSN: 0907-4449
PURE UUID: 432b09f2-20df-4e52-8e50-65c185016bc8

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Date deposited: 06 Aug 2008
Last modified: 17 Jul 2017 14:30

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Contributors

Author: V. Granata
Author: N.G. Housden
Author: S. Harrison
Author: C. Jolivet-Reynaud
Author: M.G. Gore
Author: E.A. Stura

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