title: The role of covalent dimerization on the physical and chemical stability of the EC1 domain of human E-cadherin.
abstract: The objective of this work was to evaluate the solution stability of the EC1 domain of E-cadherin under various conditions. The EC1 domain was incubated at various temperatures (4, 37, and 70 degrees C) and pH values (3.0, 7.0, and 9.0). At pH 9.0 and 37 or 70 degrees C, a significant loss of EC1 was observed due to precipitation and a hydrolysis reaction. The degradation was suppressed upon addition of dithiothreitol (DTT), suggesting that the formation of EC1 dimer facilitated the EC1 degradation. At 4 degrees C and various pH values, the EC1 secondary and tertiary showed changes upon incubation up to 28 days, and DTT prevented any structural changes upon 28 days of incubation. Molecular dynamics simulations indicated that the dimer of EC1 has higher mobility than does the monomer; this higher mobility of the EC1 dimer may contribute to instability of the EC1 domain.
authors.affiliation: Department of Pharmaceutical Chemistry, Simons Research Laboratories, The University of Kansas, 2095 Constant Ave., Lawrence, Kansas 66047.
authors.forename: Maulik
authors.initials: M
authors.lastname: Trivedi
authors.forename: R Andrew
authors.initials: RA
authors.lastname: Davis
authors.forename: Yumna
authors.initials: Y
authors.lastname: Shabaik
authors.forename: Ambrish
authors.initials: A
authors.lastname: Roy
authors.forename: Gennady
authors.initials: G
authors.lastname: Verkhivker
authors.forename: Jennifer S
authors.initials: JS
authors.lastname: Laurence
authors.forename: C Russell
authors.initials: CR
authors.lastname: Middaugh
authors.forename: Teruna J
authors.initials: TJ
authors.lastname: Siahaan
mesh_terms: D000595:Amino Acid Sequence; D015820:Cadherins / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D003198:Computer Simulation; D019281:Dimerization; D004229:Dithiothreitol / Q000032:analysis; D004355:Drug Stability; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D006863:Hydrogen-Ion Concentration; D013058:Mass Spectrometry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013050:Spectrometry, Fluorescence; D013696:Temperature
chemical_list: D015820:Cadherins; D004229:Dithiothreitol