title: Purification and secondary structural analysis of tissue inhibitor of metalloproteinases-1.
abstract: In connective tissue diseases such as rheumatoid arthritis, the matrix metalloproteinases are the primary enzymes involved in tissue degradation. Tissue inhibitor metalloproteinases-1 (TIMP-1) is a specific inhibitor of these enzymes, which is thought to regulate their action in vivo. The structure and function of TIMP-1 may therefore be important as the basis for the rational design of therapeutic agents. This paper describes a simple and effective method for the purification of sufficient quantities of TIMP-1 for spectroscopic studies. Circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy have, together, showed TIMP-1 to be mostly in a beta-sheet conformation, with significant amounts of alpha-helix and beta-turn. Two-dimensional nuclear magnetic resonance spectroscopy indicated a correspondingly high proportion of beta-sheet. CD and FTIR have also shown TIMP-1 to have high thermostability.
authors.affiliation: Rheumatology Research Unit, Addenbrooke's Hospital, Cambridge, UK.
authors.forename: D J
authors.initials: DJ
authors.lastname: Hodges
authors.forename: D C
authors.initials: DC
authors.lastname: Lee
authors.forename: C J
authors.initials: CJ
authors.lastname: Salter
authors.forename: D G
authors.initials: DG
authors.lastname: Reid
authors.forename: G P
authors.initials: GP
authors.lastname: Harper
authors.forename: T E
authors.initials: TE
authors.lastname: Cawston
mesh_terms: D002942:Circular Dichroism; D004355:Drug Stability; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification*; D006358:Hot Temperature; D009682:Magnetic Resonance Spectroscopy; D017433:Protein Structure, Secondary; D017550:Spectroscopy, Fourier Transform Infrared; D019714:Tissue Inhibitor of Metalloproteinases
chemical_list: D006023:Glycoproteins; D019714:Tissue Inhibitor of Metalloproteinases