Abstract
A combination of surface plasmon resonance (SPR) and matrix-assisted laser-desorptionionization- time-of-flight mass spectrometry (MALDI-TOF-MS) was used to study the interaction between endopolygalacturonase (PG) from Fusarium moniliforme and a polygalacturonase-inhibiting protein (PGIP) from Phaseolus vulgaris. PG hydrolyses the homogalacturonan of the plant cell wall and is considered an important pathogenicity factor of many fungi. PGIP is a specific inhibitor of fungal PGs and is thought to be involved in plant defence against phytopathogenic fungi. SPR was used either to study the effect of the PG glycosylation on the formation of the complex with PGIP, and as a sensitive affinity capture of an interacting peptide from a mixture of PG fragments obtained by limited proteolysis. Mass spectrometry allowed to characterise the interacting peptide eluted from the sensor surface.
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Selected References
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- Caprari C., Mattei B., Basile M. L., Salvi G., Crescenzi V., De Lorenzo G., Cervone F. Mutagenesis of endopolygalacturonase from Fusarium moniliforme: histidine residue 234 is critical for enzymatic and macerating activities and not for binding to polygalacturonase-inhibiting protein (PGIP). Mol Plant Microbe Interact. 1996 Sep;9(7):617–624. doi: 10.1094/mpmi-9-0617. [DOI] [PubMed] [Google Scholar]
- Gobom J., Nordhoff E., Mirgorodskaya E., Ekman R., Roepstorff P. Sample purification and preparation technique based on nano-scale reversed-phase columns for the sensitive analysis of complex peptide mixtures by matrix-assisted laser desorption/ionization mass spectrometry. J Mass Spectrom. 1999 Feb;34(2):105–116. doi: 10.1002/(SICI)1096-9888(199902)34:2<105::AID-JMS768>3.0.CO;2-4. [DOI] [PubMed] [Google Scholar]
- Ishii A., Seno H., Watanabe-Suzuki K., Kumazawa T., Matsushima H., Suzuki O., Katsumata Y. Ultrasensitive determination of phencyclidine in body fluids by surface ionization organic mass spectrometry. Anal Chem. 2000 Jan 15;72(2):404–405. doi: 10.1021/ac990765t. [DOI] [PubMed] [Google Scholar]
- Kobe B., Deisenhofer J. Mechanism of ribonuclease inhibition by ribonuclease inhibitor protein based on the crystal structure of its complex with ribonuclease A. J Mol Biol. 1996 Dec 20;264(5):1028–1043. doi: 10.1006/jmbi.1996.0694. [DOI] [PubMed] [Google Scholar]
- Leckie F., Mattei B., Capodicasa C., Hemmings A., Nuss L., Aracri B., De Lorenzo G., Cervone F. The specificity of polygalacturonase-inhibiting protein (PGIP): a single amino acid substitution in the solvent-exposed beta-strand/beta-turn region of the leucine-rich repeats (LRRs) confers a new recognition capability. EMBO J. 1999 May 4;18(9):2352–2363. doi: 10.1093/emboj/18.9.2352. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mattei B., Bernalda M. S., Federici L., Roepstorff P., Cervone F., Boffi A. Secondary structure and post-translational modifications of the leucine-rich repeat protein PGIP (polygalacturonase-inhibiting protein) from Phaseolus vulgaris. Biochemistry. 2001 Jan 16;40(2):569–576. doi: 10.1021/bi0017632. [DOI] [PubMed] [Google Scholar]
- Papageorgiou A. C., Shapiro R., Acharya K. R. Molecular recognition of human angiogenin by placental ribonuclease inhibitor--an X-ray crystallographic study at 2.0 A resolution. EMBO J. 1997 Sep 1;16(17):5162–5177. doi: 10.1093/emboj/16.17.5162. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sönksen C. P., Nordhoff E., Jansson O., Malmqvist M., Roepstorff P. Combining MALDI mass spectrometry and biomolecular interaction analysis using a biomolecular interaction analysis instrument. Anal Chem. 1998 Jul 1;70(13):2731–2736. doi: 10.1021/ac9800457. [DOI] [PubMed] [Google Scholar]
- Williams C., Addona T. A. The integration of SPR biosensors with mass spectrometry: possible applications for proteome analysis. Trends Biotechnol. 2000 Feb;18(2):45–48. doi: 10.1016/s0167-7799(99)01389-x. [DOI] [PubMed] [Google Scholar]