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Phase diagram of a crystalline protein: Determination of the solubility of concanavalin A by a microquantitation assay

Published on Sep 1, 1989in Journal of Crystal Growth1.57
· DOI :10.1016/0022-0248(89)90213-3
Vincent Mikol7
Estimated H-index: 7
(CNRS: Centre national de la recherche scientifique),
Richard Giegé53
Estimated H-index: 53
(CNRS: Centre national de la recherche scientifique)
Abstract
Abstract A quick and miniature method has been devised for determining protein solubility and used to investigate the equilibrium solubility of concanavalin A from the Jack Bean with its crystals as a function of ammonium sulfate concentration, temperature and pH. The crystals were characterized by X-ray diffraction and their morphologies related to the corresponding solubilities. The protein solution concentration was estimated out of small crystallizing drops using a rapid and sensitive microassay. Measurements of protein quantity were carried out in 96-well microplates in an automatic spectrophotometer. The resulting phase diagram has permitted to analyse the solubility of concanavalin A, to estimate supersaturation and to devise readily new ways of crystal growth of this lectin, namely by pH and temperature variations. Moreover, the approach is proved to be a valuable tool to design crystallization experiments of new molecules and to improve and control protein crystal growth.
  • References (21)
  • Citations (44)
References21
Newest
#1R. Boistelle (CNRS: Centre national de la recherche scientifique)H-Index: 20
#2Jean-Pierre Astier (CNRS: Centre national de la recherche scientifique)H-Index: 13
#1Sandra B. Howard (UAH: University of Alabama in Huntsville)H-Index: 6
#2Pamela J. Twigg (UAH: University of Alabama in Huntsville)H-Index: 3
Last.Edward J. Meehan (UAH: University of Alabama in Huntsville)H-Index: 16
view all 4 authors...
#1Keith B. Ward (NRL: United States Naval Research Laboratory)H-Index: 12
#2Mary Ann Perozzo (NRL: United States Naval Research Laboratory)H-Index: 3
Last.William M. Zuk (NRL: United States Naval Research Laboratory)H-Index: 3
view all 3 authors...
#1Naomi E. Chayen (Imperial College London)H-Index: 33
#2J. Akins (Imperial College London)H-Index: 1
Last.David M. Blow (Imperial College London)H-Index: 30
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Cited By44
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#1Joseph R. Luft (Hauptman-Woodward Medical Research Institute)H-Index: 28
#2Janet Newman (CSIRO: Commonwealth Scientific and Industrial Research Organisation)H-Index: 26
Last.Edward H. Snell (UB: University at Buffalo)H-Index: 23
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#1Kedar S. Deshpande (TU Delft: Delft University of Technology)H-Index: 3
#2Tangir Ahamed (TU Delft: Delft University of Technology)H-Index: 9
Last.Marcel Ottens (TU Delft: Delft University of Technology)H-Index: 20
view all 6 authors...
#1Alexander E. S. Van Driessche (CSIC: Spanish National Research Council)H-Index: 15
#2Jose A. Gavira (CSIC: Spanish National Research Council)H-Index: 23
Last.Fermín Otálora (CSIC: Spanish National Research Council)H-Index: 22
view all 4 authors...
#1Matthew J. Jones (MLU: Martin Luther University of Halle-Wittenberg)H-Index: 10
#2Joachim Ulrich (MLU: Martin Luther University of Halle-Wittenberg)H-Index: 25
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