Determination of pathogen-related enzyme action by mass spectrometry analysis of pectin breakdown products of plant cell walls

Hyun Joo An, Susan Lurie, L. Carl Greve, Danielle Rosenquist, Crystal Kirmiz, John M. Labavitch, Carlito B Lebrilla

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

An analytical approach using matrix-assisted laser desorption/ionization mass spectrometry for the structural characterization and assessment of the degree of polymerization of cell wall pectin-derived oligosaccharides (PDOs) in three regions of Botrytis cinerea-infected tomato fruit tissue is described. The PDOs were isolated from lesion centers (extensively macerated tissue), the area just beyond visible lesion margins, and healthy and intact tissue of an inoculated fruit, sampled at a distance from developing lesions. PDO mixtures were directly analyzed by mass spectrometry without chromatographic separation, after minimum cleanup by membrane drop dialysis. The structures identified implied the action of three different pathogen pectin-modifying enzymes. Modifications such as methyl esterification were identified by determination of exact PDO molecular masses and tandem mass spectrometry via collision-induced dissociation. We have identified four PDO series that were generated through the breakdown of homogalacturonan pectins. The decayed and lesion edge areas had fewer and less diverse PDOs than healthy tissues, possibly due to metabolic by-products of the pathogen. This analytical technique provides a simple and rapid method to characterize the pectin-derived oligosaccharides produced by in vivo digestion during pathogen infection.

Original languageEnglish (US)
Pages (from-to)71-82
Number of pages12
JournalAnalytical Biochemistry
Volume338
Issue number1
DOIs
StatePublished - Mar 1 2005

Fingerprint

Plant Cells
Pathogens
Cell Wall
Mass spectrometry
Oligosaccharides
Mass Spectrometry
Enzymes
Tissue
Fruits
Fruit
Pectins
Botrytis
Dialysis
pectin
Esterification
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Molecular mass
Lycopersicon esculentum
Tandem Mass Spectrometry
Polymerization

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

An, H. J., Lurie, S., Greve, L. C., Rosenquist, D., Kirmiz, C., Labavitch, J. M., & Lebrilla, C. B. (2005). Determination of pathogen-related enzyme action by mass spectrometry analysis of pectin breakdown products of plant cell walls. Analytical Biochemistry, 338(1), 71-82. https://doi.org/10.1016/j.ab.2004.11.004

Determination of pathogen-related enzyme action by mass spectrometry analysis of pectin breakdown products of plant cell walls. / An, Hyun Joo; Lurie, Susan; Greve, L. Carl; Rosenquist, Danielle; Kirmiz, Crystal; Labavitch, John M.; Lebrilla, Carlito B.

In: Analytical Biochemistry, Vol. 338, No. 1, 01.03.2005, p. 71-82.

Research output: Contribution to journalArticle

An, Hyun Joo ; Lurie, Susan ; Greve, L. Carl ; Rosenquist, Danielle ; Kirmiz, Crystal ; Labavitch, John M. ; Lebrilla, Carlito B. / Determination of pathogen-related enzyme action by mass spectrometry analysis of pectin breakdown products of plant cell walls. In: Analytical Biochemistry. 2005 ; Vol. 338, No. 1. pp. 71-82.
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