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@ARTICLE{Geerds:646220,
      author       = {Geerds, Christina and Niemann, Hartmut},
      title        = {{S}ingle mutations to tyrosine or glutamate improve the
                      crystallizability and crystal diffraction properties of a
                      flexible two-domain protein},
      journal      = {Acta crystallographica / Section F},
      volume       = {82},
      number       = {1},
      issn         = {1744-3091},
      address      = {Oxford [u.a.]},
      publisher    = {Blackwell},
      reportid     = {PUBDB-2026-00765},
      pages        = {4 - 13},
      year         = {2026},
      abstract     = {This case report describes single surface substitutions
                      that improve the crystallizability and diffraction
                      properties of a flexible two-domain protein. InlB392
                      comprises the internalin domain and the B repeat of the
                      Listeria monocytogenes invasion protein InlB. The InlB392
                      wild type yielded very few poorly reproducible hits in
                      crystallization screens and the crystals had a diffraction
                      limit of worse than 3.0 Å. It seems reasonable to assume
                      that this crystallization bottleneck is caused by
                      interdomain flexibility, given that crystals of the isolated
                      internalin domain or B repeat diffract to high resolution. A
                      previously identified variant, T332E, showed improved
                      crystallization and diffraction. Here, two additional
                      InlB392 variants are described with single
                      threonine-to-tyrosine or valine-to-glutamate substitutions
                      that produced crystals directly in initial screens and,
                      without optimization, diffracted to 1.6 and 1.45 Å
                      resolution, respectively. The mutated residues do not
                      participate in intramolecular interdomain interactions but
                      mediate crystal contacts, indicating that specific surface
                      properties, rather than interdomain flexibility per se,
                      impede the crystallization of wild-type InlB392. Notably,
                      the beneficial glutamate substitutions contrast with the
                      generally recognized underrepresentation of glutamate in
                      crystal contacts and the high entropic cost of fixing an
                      otherwise flexible side chain with many rotatable bonds in a
                      crystal contact. The reported results suggest that surface
                      mutations can help crystallization even if they increase the
                      entropy of the respective residue. More broadly, the
                      observations are consistent with the hypothesis that
                      negative evolutionary design limits fortuitous lattice
                      formation of proteins and the resulting expectation that
                      random mutations of surface residues are likely to improve
                      crystallizability.},
      cin          = {EMBL-User},
      ddc          = {530},
      cid          = {I:(DE-H253)EMBL-User-20120814},
      pnm          = {6G3 - PETRA III (DESY) (POF4-6G3)},
      pid          = {G:(DE-HGF)POF4-6G3},
      experiment   = {EXP:(DE-H253)P-P13-20150101},
      typ          = {PUB:(DE-HGF)16},
      doi          = {10.1107/S2053230X25010416},
      url          = {https://bib-pubdb1.desy.de/record/646220},
}