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 language | English (US) |
|---|---|
| Pages (from-to) | 71-82 |
| Number of pages | 12 |
| Journal | Analytical Biochemistry |
| Volume | 338 |
| Issue number | 1 |
| DOIs | |
| State | Published - Mar 1 2005 |
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ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology
Cite this
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 journal › Article
}
TY - JOUR
T1 - Determination of pathogen-related enzyme action by mass spectrometry analysis of pectin breakdown products of plant cell walls
AU - An, Hyun Joo
AU - Lurie, Susan
AU - Greve, L. Carl
AU - Rosenquist, Danielle
AU - Kirmiz, Crystal
AU - Labavitch, John M.
AU - Lebrilla, Carlito B
PY - 2005/3/1
Y1 - 2005/3/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=13644262157&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=13644262157&partnerID=8YFLogxK
U2 - 10.1016/j.ab.2004.11.004
DO - 10.1016/j.ab.2004.11.004
M3 - Article
C2 - 15707937
AN - SCOPUS:13644262157
VL - 338
SP - 71
EP - 82
JO - Analytical Biochemistry
JF - Analytical Biochemistry
SN - 0003-2697
IS - 1
ER -