The identification of allergen proteins in sugar beet (Beta vulgaris) pollen causing occupational allergy in greenhouses

Table 2 Protein identification after 2DE-separation of proteins in sugar beet pollen extract

Sample	Rank	Protein Name	Acc Nr	MW	pI	#
1	1	CALM1_PETHY Calmodulin-1 (CaM-1) – Petunia hybrida	P62199	16 763	4.1	6
2	1	CALM1_PETHY Calmodulin-1 (CaM-1) – Petunia hybrida	P62199	16 763	4.1	5
3	1	PROF_CUCME Profilin (Pollen allergen Cuc m 2) – Cucumis melo (Muskmelon)	Q5FX67	14 029	4.6	2
4	1	PROF_CUCME Profilin (Pollen allergen Cuc m 2) – Cucumis melo (Muskmelon)	Q5FX67	14 029	4.6	2
5	1	CHE1_CHEAL Pollen allergen Che a 1 – Chenopodium album (Lamb's-quarters)	Q8LGR0	18 739	5.0	2
	2	SODC1_MESCR Superoxide dismutase – Mesembryanthemum crystallinum	P93258	15 278	5.5	1
6	1	PMGI_MESCR 2,3-bisphosphoglycerate-independent phosphoglycerate mutase	Q42908	61 316	5.4	1
	2	SODC1_MESCR Superoxide dismutase – Mesembryanthemum crystallinum	P93258	15 278	5.5	1
	4	TRXH1_TOBAC Thioredoxin H-type 1 – Nicotiana tabacum	P29449	14 118	5.6	1
	5	CHE1_CHEAL Pollen allergen Che a 1 – Chenopodium album (Lamb's-quarters)	Q8LGR0	18 739	5.0	3
7	1	DYH1A_CHLRE Dynein-1-alpha heavy chain – Chlamydomonas reinhardtii	Q9SMH3	52 5420	5.3	1
8	1	CALM1_PETHY Calmodulin-1 (CaM-1) – Petunia hybrida	P62199	16 763	4.1	2

Samples were excised from 2DE (see Fig. 4A) and analyzed by LC-MS/MS. Protein identification after LC-MS/MS was performed with the software GPS Explorer and an in-house Mascot search engine. Settings: Precursor Tol: 15 ppm, MS/MS Fragment Tol: 0.15 Da. For each protein identification, columns show: Accession number in the Swiss-Prot database (Acc Nr), Protein theoretical mass in Da (MW), Protein theoretical isoelectric point (pI), and the number of peptides used for protein identification (#). The Mascot Best ion score (i.e. the highest score of a single peptide), and the significance of the database search (C.I. = the confidence interval) calculated by the GPS Explorer software were > 45 with at a confidence interval (C.I.) > 99.0 for all peptides except for Che a 1 in sample 6 where Best ion score was 17 and C.I. = 90.1%. Best ion score was 56 and C.I. = 99.999% for the Che a 1-homologue in sample 5. Best ion core was 82 and 62 with C.I. = 100% for the Che a 2-homologue in sample 3 and 4.

ISSN: 1476-7961