Coprecipitation of proteins with matrix ligands: scaleable protein isolation.
Matrix ligands are agents for isolating proteins out of dilute crudes by coprecipitating proteins. The ligands have a strong anion sulfonate head which initiates binding to proteins having a positive net charge, ZH+ approximately 5-20. Initial binding tightens protein conformation and starts to squeeze water from conformationally motile proteins. The tails are stackable hydrophobic organic groups, azoaromatic dyes which draw protein-ligand complexes together. Proteins coprecipitate as guests, in the ligand host matrix. In addition to stacking, ligand tails displace water because of their bulk, and lower the average dielectric constant near charged groups, which reinforces the electrostatic component of binding. Matrix ligands protect proteins, scavenge them from dilute crudes (0.01-0.1 per cent protein), and densify coprecipitates. Detergent ions in low concentrations, 10(-4)-10(-5) M also sometimes serve as coprecipitating agents, entangling their tails but probably not stacking. Divalent metal ions, Zn++, sometimes are useful auxiliary agents. Preparative scaleability from crudes is demonstrated starting from 100-200 g of raw peanuts and raw pineapple to coprecipitate a lectin and bromelain enzyme respectively in 1-2 h with 80-90 per cent activity yields. Ligands are released from coprecipitates by shifting the pH and trapping the ligands with exchange resins. Protein conformation tightening in solution is seen by viscosity measurements.
Matulis
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Biochemistry Department, University of Minnesota, St Paul 55108, USA.
Lovrien
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Richardson
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D010450:Endopeptidases / Q000302:isolation & purification; D037102:Lectins / Q000302:isolation & purification; D008024:Ligands; D015394:Molecular Structure; D019887:Peanut Agglutinin; D011506:Proteins / Q000302:isolation & purification*; D012996:Solutions / Q000737:chemistry; D013329:Structure-Activity Relationship; D015032:Zinc / Q000737:chemistry
D037102:Lectins; D008024:Ligands; D019887:Peanut Agglutinin; D011506:Proteins; D012996:Solutions; D010450:Endopeptidases; D015032:Zinc