Effect of polyethylene glycol-400 at low concentrations on long-term growth of muscle phosphoglucomutase crystals from concentrated salt solutions.
Although rabbit muscle phosphoglucomutase occasionally deposits tetragonal crystals from solutions of ammonium sulfate at about 47% of saturation, low concentrations of polyethylene glycol-400 (PEG), 1 to 4.5% w/v, must be included to sustain crystal growth. A comparison of long-term growth rates for macroscopic crystals in the presence and absence of added PEG suggests that at high salt concentration this cosolute exerts its primary effect on disordered protein aggregates, either in the external medium or at the surface of the crystal, and thereby allows the growth of much larger crystals. Since the observed effects may arise from a PEG-induced increase in the "solubility" of the aggregate that exceeds the induced increase in solubility of the crystalline phase under these conditions, the physical basis for a cosolute-induced increase in solubility in the presence of a precipitant is considered. The applicability of such a rationale to the present system is supported by an assessment of the relative effects of polyethylene glycol and beta-octylglucoside on amorphous, salt-induced precipitates of phosphoglucomutase. PEG also produces what appears to be a differential effect on nucleation efficiency and crystal growth rate. Thus, seed crystals cannot be enlarged at a significant rate at high salt concentration without producing showers of extraneous nucleation centers when the concentration of added PEG is 3% or less. But PEG concentrations of 4.5% essentially eliminate the showering problem, ostensibly by increasing the supersaturation required for nucleation to a greater extent than that required for crystal growth. The same type of effect is observed during de novo growth. Again a solubility-based mechanism is posed. Hysteretic effects related to properties of amorphous aggregates of the protein also are described.
Ray
W J
WJ
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907.
D000645:Ammonium Sulfate; D003460:Crystallization; D005960:Glucosides / Q000737:chemistry; D009132:Muscles / Q000201:enzymology*; D010733:Phosphoglucomutase / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry*; D012996:Solutions; D013997:Time Factors
D005960:Glucosides; D012996:Solutions; C018619:octyl-beta-D-glucoside; D011092:Polyethylene Glycols; D012965:Sodium Chloride; D010733:Phosphoglucomutase; D000645:Ammonium Sulfate