BPTI liquid-liquid phase separation monitored by light and small-angle X-ray scattering

被引:33
作者
Grouazel, S
Perez, J
Astier, JP
Bonneté, F
Veesler, S
机构
[1] Univ Aix Marseille 2, CNRS, Ctr Rech Mecanismes Croissance Cristalline, F-13288 Marseille 09, France
[2] Univ Aix Marseille 3, CNRS, Ctr Rech Mecanismes Croissance Cristalline, F-13288 Marseille, France
[3] Ctr Univ Paris Sud, LURE, CNRS, CEA,MENRT, F-91898 Orsay, France
来源
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY | 2002年 / 58卷
关键词
BPTI; liquid-liquid phase separation; small-angle X-ray scattering;
D O I
10.1107/S0907444902012799
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
In the field of protein crystallization, a better knowledge of the nucleation process is essential to control the nucleation rate, the growth and therefore the size and the quality of crystals. With that aim, it becomes clear that the important stage is the determination of the protein phase diagram. We highlighted and investigated the bovine pancreatic trypsin inhibitor (BPTI) binary liquid-liquid phase separation in 350 mM KSCN solutions as a function of temperature. We measured the low concentration part of the binodal curve using light scattering and optical microscopy. We show, from small angle X-ray scattering experiments, that the high concentrated phase sediments in the bottom of the capillary and we analysed the low concentrated phase in terms of monomers/decamers equilibrium.
引用
收藏
页码:1560 / 1563
页数:4
相关论文
共 17 条
[1]   DATA APPRAISAL, EVALUATION AND DISPLAY FOR SYNCHROTRON RADIATION EXPERIMENTS - HARDWARE AND SOFTWARE [J].
BOULIN, C ;
KEMPF, R ;
KOCH, MHJ ;
MCLAUGHLIN, SM .
NUCLEAR INSTRUMENTS & METHODS IN PHYSICS RESEARCH SECTION A-ACCELERATORS SPECTROMETERS DETECTORS AND ASSOCIATED EQUIPMENT, 1986, 249 (2-3) :399-407
[2]   Investigation of aprotinin (BPTI) solutions during nucleation [J].
Budayova-Spano, M ;
Bonneté, F ;
Astier, JP ;
Veesler, S .
JOURNAL OF CRYSTAL GROWTH, 2002, 235 (1-4) :547-554
[3]  
DEPAUTEX C, 1987, LURE RAPPORT ACTIVIT
[4]   Improved signal-to-background ratio in small-angle X-ray scattering experiments with synchrotron radiation using an evacuated cell for solutions [J].
Dubuisson, JM ;
Decamps, T ;
Vachette, P .
JOURNAL OF APPLIED CRYSTALLOGRAPHY, 1997, 30 :49-54
[5]   Control of protein crystal nucleation around the metastable liquid-liquid phase boundary [J].
Galkin, O ;
Vekilov, PG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (12) :6277-6281
[6]   Understanding protein crystallization on the basis of the phase diagram [J].
Haas, C ;
Drenth, J .
JOURNAL OF CRYSTAL GROWTH, 1999, 196 (2-4) :388-394
[7]   The BPTI decamer observed in acidic pH crystal forms pre-exists as a stable species in solution [J].
Hamiaux, C ;
Pérez, J ;
Prangé, T ;
Veesler, S ;
Riès-Kautt, M ;
Vachette, P .
JOURNAL OF MOLECULAR BIOLOGY, 2000, 297 (03) :697-712
[8]   The decameric structure of bovine pancreatic trypsin inhibitor (BPTI) crystallized from thiocyanate at 2.7Å resolution [J].
Hamiaux, C ;
Prangé, T ;
Riès-Kautt, M ;
Ducruix, A ;
Lafont, S ;
Astier, JP ;
Veesler, S .
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 1999, 55 :103-113
[9]  
HAMIAUX C, 2000, THESIS U PARIS 11
[10]   Comparison of solubilities and molecular interactions of BPTI molecules giving different polymorphs [J].
Lafont, S ;
Veesler, S ;
Astier, JP ;
Boistelle, R .
JOURNAL OF CRYSTAL GROWTH, 1997, 173 (1-2) :132-140