Non‐ionic Adsorptive Immobilization of Proteins to Palmityl‐Substituted Sepharose 4B
Open Access
- 1 April 1982
- journal article
- research article
- Published by Wiley in European Journal of Biochemistry
- Vol. 123 (3), 601-610
- https://doi.org/10.1111/j.1432-1033.1982.tb06575.x
Abstract
1 Palmityl-substituted Sepharose 4B prepared by the glycidyl either method of S. Hjertén et al. [J. Chromatogr. (1974) 101, 281–288] has been used as a non-ionic matrix for protein adsorption. A number of proteins, some of which are catalytically active, may be immobilized on this adsorbent in the form of suspension or column. 2 Of the proteins examined, bovine serum albumin, hemoglobin, myoglobin, lysozyme, glutamate dehydrogenase, and β-galactosidase were immobilized on the adsorbent irrespective of NaCl concentration. Trypsin, α-chymotrypsin, papain, pepsin, and amyloglucosidase were totally adsorbed either in the absence of any additional salt or at high salt concentrations. Cytochrome c, used as a model protein, was totally immobilized only ar high ionic strength and low pH. 3 Immobilization normaly took place with an apparent in initial acitivity rates. In the case of trypsin using Nα-benzoyl-cl-arginine p-nitroanilide as substrate, adsorption resulted in anincrease in V (app). 4 Beef-liver glutamate dehydrogenase, used as a model allosteric enzyme, was found to retain its allosteric properties towards ADP and GTP after immobilization. 5 Results are discussed in terms of specific interactions involving a smaller number of binding sites in protein molecules as compared to the multiple attachment to highly substituted adsorbents prepared with shorted ligands. Retention of the essential properties of the proteins examined in this study are ascribed to these characteristics of the adsorbent and to its non-ionic nature. Relevance of these observations to in vivo processes and the potential use of the adsorbent for enzyme immobilization are also discussed.This publication has 59 references indexed in Scilit:
- Accessible hydrophobic groups of native proteinsBiochemical and Biophysical Research Communications, 1975
- Protein binding by agarose carrying hydrophobic groups in conjunction with chargesBiochemical and Biophysical Research Communications, 1973
- Hydrocarbon-coated Sepharoses. Use in the purification of glycogen phosphorylaseBiochemical and Biophysical Research Communications, 1972
- Regulatory effects of mitochondrial lipids on glutamate dehydrogenase (NAD(P))FEBS Letters, 1972
- Effects of neutralization of amino groups on the kinetics of chymotrypsinBiochemical and Biophysical Research Communications, 1970
- Chemical Coupling of Peptides and Proteins to Polysaccharides by Means of Cyanogen HalidesNature, 1967
- The Statistical Analysis of Enzyme Kinetic DataPublished by Wiley ,1967
- Contribution of Hydrophobic Interactions to the Stability of the Globular Conformation of ProteinsJournal of the American Chemical Society, 1962
- Intrinsic Viscosity and Optical Rotation of Proteins in Acid Media1Journal of the American Chemical Society, 1955
- Reduction of Organic Compounds by Lithium Aluminum Hydride. II. Carboxylic AcidsJournal of the American Chemical Society, 1947