Nucleation and Growth Kinetics of Protein Crystals

Results obtained by this project were summarized as below.(1)Japanese team developed a Michelson Interferometer objective lens of super-long working distance(25.5毫米)with Nikon Co.Using this interferometer，a novel technique for determining solubility was developed，and solubilities of tetragonal and orthorhombic lysozyme crystal were detemined as a function of temperature for the first time.(2)netragonal and orthorthorhombic lysozyme crystal were detemined as s.Equipment to measure magnetic birefringence was developed.(3)Japanese team highly-purified commercial lysozyme(99.9%)by purification technique developed by U.S.A.team.Growthprocess of lysozyme crystals were in situ observed by atomic force microscopy.It was found that small amount of impurity(1%)drastically decreased growth rate of(101)face of tetragonal crystal.(4)Japanese team developed a novel technique to measure impurity content and its distribution in the crystal with a fluorescense indicator.A novel mechanism of the impurity incorporation into the crystal was proposed.(5)U.S.A.team analyzed the concentration distribution around growing lysozyme crystals using computer simulation technique.Japanese team measured the actual concentration distribution around the crystal by interferometry.These two results were compared each other and found that they mostly agreed.It was found that buoyancy driven convection can be stopped when the thickness of the solution is smaller than l50o m.(6)Japanese team observed the concentration distribution around lysozyme crystals under lg to micro-g using free fall facilities。The changes in the concentration distribution were analyzed.

Results obtained by this project were summarized as below.(1) Japanese team developed a Michelson Interferometer objective lens of super-long working distance (25.5 mm) with Nikon Co. Using this interferometer, a novel technique for determining solubility was developed, and solubilities of tetragonal and orthorhombic lysozyme crystal were detemined as a function of temperature for the first time.(2) Japanese team studied magnetic field effects on the crystallization of lysozyme, and found that a magnetic field (i) increased a growth rate and (ii) decreased a solubility. Equipment to measure magnetic birefringence was developed.(3) Japanese team highly-purified commercial lysozyme (99.9%) by purification technique developed by U.S.A.team. Growthprocess of lysozyme crystals were in situ observed by atomic force microscopy. It was found that small amount of impurity (1%) drastically decreased growth rate of (101) face of tetragonal crystal.(4) Japanese team developed a novel technique to measure impurity content and its distribution in the crystal with a fluorescense indicator. A novel mechanism of the impurity incorporation into the crystal was proposed.(5) U.S.A.team analyzed the concentration distribution around growing lysozyme crystals using computer simulation technique. Japanese team measured the actual concentration distribution around the crystal by interferometry. These two results were compared each other and found that they mostly agreed. It was found that buoyancy driven convection can be stopped when the thickness of the solution is smaller than l50オm.(6) Japanese team observed the concentration distribution around lysozyme crystals under lg to micro-g using free fall facilities. The changes in the concentration distribution were analyzed.