title: Stabilization of proteins by low molecular weight multi-ions.
abstract: A method is described to identify small molecule ligands that stabilize proteins. The procedure is based on the hypothesis that molecules of various sizes containing two to four charges should occasionally bind to unpaired charged sites on the surface of proteins and by crosslinking such residues stabilize the native state of the liganded protein. A simple turbidity assay is employed that detects inhibition of protein aggregation under selected sets of conditions. Eight test proteins were screened and in all cases specific ligands were identified that inhibited protein aggregation at millimolar to micromolar concentrations. Only small effects of these stabilizers on protein biological activities were found. In some, but not all cases, circular dichroism and fluorescence studies provided direct evidence of the binding of stabilizing ligands to the proteins suggesting multiple mechanisms of stabilization. This approach should be applicable to the development of excipients for the stabilization of pharmaceutical proteins and industrial enzymes as well as serve as starting points for second-generation inhibitors of increased affinity and specificity.
authors.affiliation: Department of Pharmaceutical Chemistry, University of Kansas, 2095 Constant Avenue, Lawrence, Kansas 66047, USA.
authors.forename: Donald S
authors.initials: DS
authors.lastname: Maclean
authors.forename: Quansheng
authors.initials: Q
authors.lastname: Qian
authors.forename: C Russell
authors.initials: CR
authors.lastname: Middaugh
mesh_terms: D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002942:Circular Dichroism; D004353:Drug Evaluation, Preclinical; D004355:Drug Stability; D004563:Electrochemistry; D007477:Ions; D007700:Kinetics; D008970:Molecular Weight; D009391:Nephelometry and Turbidimetry; D010084:Oxidation-Reduction; D011506:Proteins / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature
chemical_list: D007477:Ions; D011506:Proteins