Paper presented at the 23rd Informal Meeting on Mass Spectrometry, Fiera di Primiero, Italy, 15–19 May 2005.
Activity of anchored human matrix metalloproteinase-1 catalytic domain on Au (111) surfaces monitored by ESI-MS†
Version of Record online: 30 NOV 2005
Copyright © 2005 John Wiley & Sons, Ltd.
Journal of Mass Spectrometry
Volume 40, Issue 12, pages 1565–1571, December 2005
How to Cite
Grasso, G., D'Agata, R., Rizzarelli, E., Spoto, G., D'Andrea, L., Pedone, C., Picardi, A., Romanelli, A., Fragai, M. and Yeo, K. J. (2005), Activity of anchored human matrix metalloproteinase-1 catalytic domain on Au (111) surfaces monitored by ESI-MS. J. Mass Spectrom., 40: 1565–1571. doi: 10.1002/jms.929
- Issue online: 30 NOV 2005
- Version of Record online: 30 NOV 2005
- Manuscript Accepted: 11 AUG 2005
- Manuscript Received: 24 MAY 2005
- MIUR. Grant Numbers: (FIRB 2003 RBNE01TTJW004, PRIN 2003 n 2003091372).
- matrix metalloproteinases;
- human fibroblast collagenase;
- activity assay;
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. Copyright © 2005 John Wiley & Sons, Ltd.