title: Activity of anchored human matrix metalloproteinase-1 catalytic domain on Au (111) surfaces monitored by ESI-MS.
abstract: Matrix metalloproteinases (MMPs) are a family of Zn-dependent endo-peptidases known for their ability to cleave several components of the extracellular matrix, but which can also cleave many non-matrix proteins. There are many evidences that MMPs are involved in physiological and pathological processes, and a huge effort has been put in the development of possible inhibitors that could reduce the activity of MMPs, as it is clear that the ability to monitor and control such activity plays a pivotal role in the search for potential drugs aimed at finding a cure for several diseases such as pulmonary emphysema, rheumatoid arthritis, fibrotic disorders and cancer.A powerful method currently available to study enzyme-inhibitor interactions is based on the use of the surface plasmon resonance (SPR) technique. When MMP interactions are studied, a procedure by which inhibitors are normally anchored on sensor chips and SPR technique is used in order to study their interaction with MMPs molecules is usually followed. This is because it is currently believed that MMPs cannot be anchored on the sensor-chip surface without losing their activity. However, this approach gives rise to problems, as the anchoring of low-molecular-weight inhibitors on gold surfaces easily affects their ability to interact with MMPs. For this reason, the anchoring of MMPs is highly desirable.A new experimental protocol that couples the Fourier transform-SPR (FT-SPR) technique with electrospray ionization-mass spectroscopy (ESI-MS) is described here for the evaluation of the activity of MMP-1 catalytic domain (cdMMP-1) anchored on gold surfaces. The cdMMP-1 surface coverage is calculated by using FT-SPR and the enzyme activity is estimated by ESI-MS. The proposed method is label-free.
authors.affiliation: Consorzio Interuniversitario di Ricerca in Chimica dei Metalli nei Sistemi Biologici, Via C. Ulpiani 27, 70126, Bari, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Grasso
authors.forename: Roberta
authors.initials: R
authors.lastname: D'Agata
authors.forename: Enrico
authors.initials: E
authors.lastname: Rizzarelli
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Spoto
authors.forename: Luca
authors.initials: L
authors.lastname: D'Andrea
authors.forename: Carlo
authors.initials: C
authors.lastname: Pedone
authors.forename: Andrea
authors.initials: A
authors.lastname: Picardi
authors.forename: Alessandra
authors.initials: A
authors.lastname: Romanelli
authors.forename: Marco
authors.initials: M
authors.lastname: Fragai
authors.forename: Kwon Joo
authors.initials: KJ
authors.lastname: Yeo
mesh_terms: D020134:Catalytic Domain / Q000502:physiology*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005583:Fourier Analysis; D006046:Gold / Q000737:chemistry*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D007700:Kinetics; D020781:Matrix Metalloproteinase 1 / Q000737:chemistry* / Q000378:metabolism*; D061965:Matrix Metalloproteinase Inhibitors; D009842:Oligopeptides / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013449:Sulfonamides / Q000494:pharmacology; D020349:Surface Plasmon Resonance
chemical_list: D004800:Enzymes, Immobilized; D006877:Hydroxamic Acids; D061965:Matrix Metalloproteinase Inhibitors; C500716:N-isobutyl-N-(4-methoxyphenylsulfonyl)glycylhydroxamic acid; D009842:Oligopeptides; D013449:Sulfonamides; D006046:Gold; D020781:Matrix Metalloproteinase 1