@article{MishraSecklerBhat2005,
  author    = {Mishra, Rajesh and Seckler, Robert and Bhat, Rajiv},
  title     = {Efficient refolding of aggregation-prone citrate synthase by polyol osmolytes : how well are protein folding and stability aspects coupled?},
  issn      = {0021-9258},
  year      = {2005},
  abstract  = {Efficient refolding of proteins and prevention of their aggregation during folding are of vital importance in recombinant protein production and in finding cures for several diseases. We have used citrate synthase ( CS) as a model to understand the mechanism of aggregation during refolding and its prevention using several known structure-stabilizing cosolvent additives of the polyol series. Interestingly, no parallel correlation between the folding effect and the general stabilizing effect exerted by polyols was observed. Although increasing concentrations of polyols increased protein stability in general, the refolding yields for CS decreased at higher polyol concentrations, with erythritol reducing the folding yields at all concentrations tested. Among the various polyols used, glycerol was the most effective in enhancing the CS refolding yield, and a complete recovery of enzymatic activity was obtained at 7 M glycerol and 10 mu g/ml protein, a result superior to the action of the molecular chaperones GroEL and GroES in vitro. A good correlation between the refolding yields and the suppression of protein aggregation by glycerol was observed, with no aggregation detected at 7 M. The polyols prevented the aggregation of CS depending on the number of hydroxyl groups in them. Stopped-flow fluorescence kinetics experiments suggested that polyols, including glycerol, act very early in the refolding process, as no fast and slow phases were detectable. The results conclusively demonstrate that both the thermodynamic and kinetic aspects are critical in the folding process and that all structure-stabilizing molecules need not always help in productive folding to the native state. These findings are important for the rational design of small molecules for efficient refolding of various aggregation-prone proteins of commercial and medical relevance},
  language  = {en}
}
@article{SecklerLilie2005,
  author    = {Seckler, Robert and Lilie, Hauke},
  title     = {Folding and association of multi-domain and oligomeric proteins},
  isbn      = {978- 3-527-30784-5},
  year      = {2005},
  language  = {en}
}
@article{KaufmannBaxaChipmanetal.2005,
  author    = {Kaufmann, B. and Baxa, Ulrich and Chipman, P. R. and Rossmann, M. G. and Modrow, Susanne and Seckler, Robert},
  title     = {Parvovirus B19 does not bind to membrane-associated globoside in vitro},
  issn      = {0042-6822},
  year      = {2005},
  abstract  = {The glycosphingolipid globoside (globotetraosylceramide, Gb4Cer) has been proposed to be the cellular receptor of human parvovirus B19. Quantitative measurements of the binding of parvovirus B19 to Gb4Cer were performed to explore the molecular basis of the virus tropism. Solid-phase assays with fluorescence-labeled liposomes or (125)iodine-labeled empty capsids were used to characterize the specificity of binding. In addition, surface plasmon resonance on lipid layers, as well as isothermal titration microcalorimetry, was utilized for real-time analysis of the virus-receptor interaction. These studies did not confirm binding of Gb4Cer to recombinant B19 VP2 capsids, suggesting that Gb4Cer does not function on its own as the cellular receptor of human parvovirus B19, but might be involved in a more complex recognition event. The biochemical results were further confirmed by cryo-electron microscopy image reconstructions at 10 A resolution, in which the structures of empty capsids were compared with empty capsids incubated with Gb4Cer. (C) 2004 Elsevier Inc. All rights reserved},
  language  = {en}
}