Annex Kleter & Peijnenburg
Potential linear IgE-epitopes in novel proteins
Appendix - Identical sequences (minimal 6 amino acids)
shared by novel proteins and allergens
Supplementary material provided in appendix to “Presence of potential allergy-related linear
epitopes in novel proteins from conventional crops and the implication for the safety
assessment of these crops with respect to the current testing of genetically modified crops”
by Gijs A. Kleter and Ad A.C.M. Peijnenburg, submitted to Plant Biotechnology Journal.
Method
The same methodology has been applied as previously described [Kleter and Peijnenburg,
2002].
Sequences of novel proteins
Novel protein sequences are obtained from protein databanks (e.g. Entrez Protein:
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=Protein) or from literature. Alternative
protein databases that may be consulted are:

EMBL, http://srs.embl-heidelberg.de:8000/srs5/

PIR-NREF, http://pir.georgetown.edu/

SwissProt, http://www.expasy.ch
Comparison of sequences
To search for identical sequences of at least six contiguous amino acids between the
sequences of selected novel proteins and those of known allergenic proteins:

Select the Protein BLAST algorithm to “search for short nearly exact matches” from the
BLAST webpage of the U.S. National Library of Medicine’s Entrez website:
http://www.ncbi.nlm.nih.gov/BLAST/

The following default parameters are used:
expect: 20000; word size: 2; matrix: PAM 30; gap costs: existence 9, extension 1
The following parameters are user defined:
limit by Entrez query: allergen; descriptions: 500; alignments: 500
Note: in our experience, changing the default values does not improve the outcome in
such a way that sequence identities of six or more contiguous amino acids could be
more easily selected from the list. The user defined criteria have been in our
experience sufficient to obtain a list containing all identical stretches of six or more
amino acids that match proteins retrieved by the query limit “allergen”.

Enter the sequence of the novel protein into the appropriate window and click the
“BLAST” button and the “Format” button in the webpage that subsequently appears.

From this webpage, select the identical peptide sequences of at least six contiguous
amino acids.

Access the data files on the proteins with identical stretches through the hyperlinks in
the BLAST result list. Exclude from the results those proteins that are not allergens but
that have been retrieved by the query limit “allergen” because they carry the term
“allergen” in their filetext, such as proteins that belong to protein families that contain
allergens as well as antibodies reacting with allergens.
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Annex Kleter & Peijnenburg
Potential linear IgE-epitopes in novel proteins
Antigenicity prediction
Carry out antigenicity prediction by constructing a hydrophilicity plot with the Hopp and
Woods method [Hopp and Woods, 1981]:

Access the pertinent webpage on the Colorado State University’s website:
http://arbl.cvmbs.colostate.edu/molkit/hydropathy/index.html
The following webpages can be accessed alternatively for this purpose:

Expasy: http://us.expasy.org/cgi-bin/protscale.pl (window size 6 amino acids not
available)

Weizmann Institute: http://bioinformatics.weizmann.ac.il/hydroph/

Enter the protein sequence in the box and select the following settings:
Hopp & Woods, Window size: 6
Choose “plot”

In the plot, check whether the top of the highest peak coincides with the identical
stretch identified by the BLAST search within the protein sequence. The presence of
this top within the stretch would indicate that it has a high probability of antibodybinding (antigenicity).
Literature research
Search the scientific literature for data on IgE-binding epitopes within the allergenic proteins
that contain the identical stretches that have been identified in the sequence comparison.
Check if linear IgE-binding epitopes contain the identical stretch. To search literature,
PubMed, an on-line biomedical bibliography, can be used
(http://www.ncbi.nlm.nih.gov/entrez/query.fcgi). In addition, a number of allergen databases
on the Internet provide information on allergenic proteins and, occasionally, linear IgEepitopes. Examples of Internet-accessible databases on allergens and epitopes are:

CSL (free registration required): http://www.csl.gov.uk/allergen

Protall: http://www.ifr.bbsrc.ac.uk/protall

Allergome (free registration required): http://www.allergome.org

SDAP: http://129.109.73.75/SDAP/
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Annex Kleter & Peijnenburg
Potential linear IgE-epitopes in novel proteins
Results
*=
aa =
gi =
allergenic protein listed by IUIS/WHO (http://www.allergen.org)
amino acid
general identifier for protein accession file (retrievable from protein databank on NCBI
website; http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=Protein)
Antigenic? (prediction):
“yes” = the highest scoring point of the antigenicity plot falls within the identical sequence
“-“ =
the highest scoring point does not fall within the identical sequence.
Prediction method: Hopp and Woods [1981], window size = six amino acids.
1. ORFA501, Allium schoenophrasum (chives), CMS1 cytoplasm
Length:
167 aa
Accession:
not applicable
Reference DNA sequence:
Engelke and Tatioglu [2002]
RNA editing:
not applicable
Results:
no stretches of at least 6 aa identical with allergens
2. ORF138, Brassica napus (oilseed rape) and Raphanus sativus (radish), ogu
cytoplasm
Length:
138 aa
Accession:
gi 732177
Reference amino acid sequence:
Bonhomme et al. [1992], Krishnasamy and Makaroff
[1993]
RNA editing:
yes, edited sequences E, F, and H have also been
used for BLAST searches (1)
Results:
1 identical stretch, see table
Identical sequence
Allergen (2)
name
KEEKKE (3)
(1)
(2)
(3)
(4)
(5)
(6)
(7)
Phl p 13 * (4)
length
aa
394
source
Phleum pratense, grass pollen
Antigenic?
(prediction)
novel
allerprotein
gen
Yes (6)
Yes (5)
Part of linear IgE
epitope?
Yes (7)
Amino acid sequences after RNA editing in cultivated radishes [Yamagashi and Terachi, 2001]:
edit B:
same as original sequence
edit E:
T(aa3)  P(aa3); F(aa32)  Y(aa32)
edit F:
Same as E, with deletion of C-terminal 13 aa (total of 125 aa remaining)
edit H: T(aa3)  P(aa3); K(aa21)  Q(aa21); L(aa30)  F(aa30); F(aa32)  Y(aa32); I(aa110)  K(aa110)
Accessions:
Phl p 13: gi 4826572
Sequence KEEKKE is present in threefold in ORF138 and edited ORF138 B, E, and H, and in twofold in edited ORF138 F
and polygalacturonase (Phleum pratense). The two KEEKKE sequences in polygalacturonase are partially overlapping
Denoted by accession file as polygalacturonase
Each of the KEEKKE sequences of the novel protein contains one peak with the same height in the antigenicity plot; the
peaks of the KEEKKE sequences are the highest in the plot
Both overlapping KEEKKE sequences of Phl p 13 are part of the same highest plateau within the antigenicity plot
Sequence KEEKKE corresponds to the KXEE/KEXE (X = empty, T, or A) IgE-binding motif sequence of latex allergen Hev
b 5 in immunised mice [Slater et al., 1999]
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Annex Kleter & Peijnenburg
Potential linear IgE-epitopes in novel proteins
3. ORF222, Brassica napus (oilseed rape), nap cytoplasm
Length:
222 aa
Accession:
gi 1800191
Reference amino acid sequence:
L’Homme et al. [1997]
RNA editing:
not applicable
Results:
1 identical sequence, see table
Identical sequence
Allergen (1)
name
SKLKLG
(1)
(2)
Der f 15 (2)
length
aa
555
source
Dermatophagoides farinae
Antigenic?
(prediction)
novel
allerprotein
gen
-
Part of linear IgE
epitope?
Accessions:
Der f 15: gi 5815436
Denoted by accession file as 98kDa HDM allergen
4. ORF224, Brassica napus (oilseed rape), pol cytoplasm
Length:
224 aa
Accession:
gi 257560
Reference amino acid sequence:
Singh and Brown [1991]
RNA editing:
yes, an edited sequence has also been used for
BLAST searches (1)
Results:
no stretches of at least 6 aa identical with allergens
(1)
Amino acid sequences after RNA editing [Handa et al., 1995; Stahl et al., 1994]:
R(aa71)  W(aa71)
5. ORFB, Daucus carota (carrot)
Length:
Accession:
Reference amino acid sequence:
RNA editing:
Results:
(1)
(2)
216 aa
gi 10801544 (1)
Nakajima et al. [2001]
yes, edited sequences have also been used for
BLAST searches (2)
no stretches of at least 6 aa identical with allergens
The literature sequence differs from the accession sequence as follows:
W(aa28, accession)  C(aa28, literature); Y(aa122, accession)  P(aa122, literature);
Both the accession- and the literature- sequences have been used for BLAST searches
Amino acid sequences after RNA editing [Nakajima et al., 2001]:
S(aa16)  L(aa16); L(aa20)  F(aa20); P(aa151)  L(aa151)
Edited versions of the literature- and accession- sequences have been used for BLAST searches
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Annex Kleter & Peijnenburg
Potential linear IgE-epitopes in novel proteins
6. ORFH522, Helianthus annuus (sunflower), PET1 cytoplasm
Length:
174 aa
Accession:
gi 12987
Reference amino acid sequence:
Koehler et al. [1991]
RNA editing:
not applicable
Results:
3 identical sequences, see table
Identical sequence
Allergen (1)
name
ISTLGA
TATAAV
TATAAV
(1)
(2)
(3)
isoflavone
reductase-like
protein CJP-6
Phl p 5 precursor *,
Phl p 5 *, Phl p 5a *,
Phl p 5.0103
precursor *
Poa p 5 precursor *,
Poa p 5 * (3)
Antigenic?
(prediction)
novel
allerprotein
gen
-
Part of linear IgE
epitope?
length
aa
306
source
276
(Phl p 5)
Timothy grass pollen, Phleum
pratense
-
-
(2)
307
(Poa p 5)
Kentucky blue grass pollen, Poa
pratensis
-
-
(2)
Cedar pollen, Cryptomeria japonica
Accessions:
Isoflavone reductase-like protein CJP-6: gi 19847822
Phl p 5 precursor: gi398830, gi 480814, gi 739542
Phl p 5: gi 1684720, gi 3135497, gi 3135499, gi 3135503
Phl p 5a: gi 2398757, gi 2851456
Phl p 5.0103 precursor: gi 3309039
Poa p 5 precursor: gi 113562, gi 169631
Poa p 5: gi 100733
Corresponding sequences AATAAV and AATAAA are part of peptides (33 and 12 aa size, respectively) derived from
analogous allergens rHol l 5 01 and Lol p 5, respectively, that are bound by antisera from grass pollen-allergic patients
[Schramm et al., 2001; Suphioglu et al., 1998]. Deletion or mutation of the AATAAA sequence from the peptides did not
abolish IgE-binding [Suphioglu et al., 1998]. The AATAAA sequence therefore appears to be non-essential for IgE-binding.
Denoted by accession files as Poa p IX
7. ORF79, Oryza sativa (rice), Bo cytoplasm
Length:
79 aa
Accession:
gi 440860
Reference amino acid sequence:
Akagi et al. [1994]
RNA editing:
not applicable
Results:
no stretches of at least 6 aa identical with allergens
8. ORF239, Phaseolus vulgaris (common bean)
Length:
239 aa
Accession:
gi 169321
Reference amino acid sequence:
Johns et al. [1992]
RNA editing:
not applicable
Results:
1 identical sequence, see table
Identical sequence
Allergen (1)
name
length
aa
111
source
Antigenic?
(prediction)
novel
allerprotein
gen
-
Part of linear IgE
epitope?
VPSGGS
Cla h 4 * (2)
Mould Cladosporium herbarum
(1) Accessions:
Cla h 4: gi 1173074, gi 452606
(2) Denoted by accession file as Cla h 3, 60S acidic ribosomal protein P2, which corresponds, however, to Cla h 4 in the
IUIS/WHO list, i.e. see also Table 1 of a recent review on mould allergens [Breitenbach and Simon Nobbe, 2002]
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Annex Kleter & Peijnenburg
Potential linear IgE-epitopes in novel proteins
9. ORF256, Triticum aestivum (wheat)
Length:
256 aa
Accession:
gi 903885
Reference amino acid sequence:
Rathburn and Hedgcoth [1991] (1)
RNA editing:
yes, an edited sequence has also been used for
BLAST searches (2)
Results:
1 identical sequence, see table
Identical sequence
Allergen (3)
name
QALAHL
(1)
(2)
(3)
Lep d 5 *
length
aa
110
source
Storage mite Lepidoglyphus
destructor
Antigenic?
(prediction)
novel
allerprotein
gen
-
Part of linear IgE
epitope?
Literature sequence appears to be the edited form of accession file sequence; both sequences have been included
Amino acid sequence after RNA editing [Rathburn and Hedgcoth, 1991]
R(aa114, accession)  W(aa114, literature)
R(aa181, accession)  W(aa181, literature)
R(aa198, accession)  W(aa198, literature)
Accessions:
Lep d 5: gi 14423651; gi 6523378
10. ORF77, Zea mays (maize), S cytoplasm
Length:
40 aa
Accession:
not applicable
Reference amino acid sequence:
Zabala et al. [1997]
RNA editing:
not applicable
Results:
no stretches of at least 6 aa identical with allergens
11. ORF355, Zea mays (maize), S cytoplasm
Length:
355 aa
Accession:
gi 2266936
Reference amino acid sequence:
Zabala et al. [1997]
RNA editing:
not applicable
Results:
2 identical sequences, see table
Identical sequence
Allergen (1)
name
LSLDKK
Der f 16 *
ASVPAA
Phl p 5b *, Phl p
5.0203 precursor,
Phl p 5.0204, Phl p
5.0206 precursor,
Phl p 5.0207
precursor
(1)
(2)
(3)
length
aa
480
284
(Phl p 5b)
source
Housedust mite Dermatophagoides
farinae
Grass pollen, Phleum pratense
Antigenic?
(prediction)
novel
allerprotein
gen
- (2)
Part of linear IgE
epitope?
-
(3)
-
Accessions:
Der f 16: gi 21591547
Phl p 5b: gi 2398759, gi 2851457
Phl p 5.0203 precursor: gi 3309041
Phl p 5.0204: gi 3309043
Phl p 5.0206 precursor: gi 3309045
Phl p 5.0207 precursor: gi 3309047
One of the two highest points of antigenicity plot of Der f 16 immediately flanks C-terminal residue of sequence ASVPAA
Sequences corresponding to ASVPAA in Lol p 5 (ANAPPA) and Phl p 5 (AVVPPA) are part of peptides that react with
antisera from grass pollen-allergic patients [Schramm et al., 2001; Suphioglu et al., 1998]. Deletion of the ANAPPA
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Annex Kleter & Peijnenburg
Potential linear IgE-epitopes in novel proteins
sequence from the peptides does not abolish IgE-binding [Suphioglu et al., 1998]. The ANAPPA sequence therefore
appears to be non-essential for IgE-binding.
12. ORF13, Zea mays (maize), T cytoplasm
Length:
115 aa
Accession:
gi 82756
Reference amino acid sequence:
Dewey et al. [1986]
RNA editing:
not applicable
Results:
no stretches of at least 6 aa identical with allergens
13. ORF107, Sorghum bicolor (sorghum), A3 cytoplasm
Length:
107 aa
Accession:
gi 7489656
Reference amino acid sequence:
Tang et al. [1996]
RNA editing:
yes, edited sequences have also been used for
BLAST searches (1)
Results:
no stretches of at least 6 aa identical with allergens
(1)
Amino acid sequences after RNA editing [Tang et al., 1996]:
L(aa12)  S(aa12); F(aa20)  S(aa20); F(aa39)  S(aa39); S(aa40)  P(aa40);
14. dsRNA, Pyrus pyrifolia (sand pear)
Length:
477 aa
Accession:
gi 3970837
Reference amino acid sequence:
Osaki et al. [1998]
RNA editing:
not applicable
Results:
1 identical sequence, see table
Identical sequence
Allergen (1)
name
SVPDVL
(1)
Fus c 2 *
length
aa
121
source
Mould Fusarium culmorum
Antigenic?
(prediction)
novel
allerprotein
gen
-
Part of linear IgE
epitope?
Accessions:
Fus c 2: gi 19879659
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Annex Kleter & Peijnenburg
Potential linear IgE-epitopes in novel proteins
15. dsRNA, Oryza sativa (rice)
Length:
Accession:
Reference amino acid sequence:
RNA editing:
Results:
Identical sequence
4572 aa
gi 1030716
Moriyama et al. [1995]
not applicable
18 identical sequences, see table
Allergen (1)
name
NLGTTA
Pen m 2 *
length
aa
356
EIEEFR
Phl p 11 *
143
EIEEFR
Lol p 11 *
134
SNLDEF
Cop c 2 *
106
QSDRLQ
Ara h 2 *, Ara h 2.02
SLSNSSF
DENEKL
Bet v 3 *
Ana o 1 (3) *
156
(Ara h 2)
205
536, 538
DENEKL
NWYYKR
Cor a 11 (5) *
Major allergen 1
448
330
VHRFNEG
SLNYDG
Ara h 4 (6) *
Equ c 1 *, Equ c 1
chain A
Cla h 5 *
Der f 16 *
530
187
(Equ c 1)
204
480
631
211
(Eur m 1)
LRTVTP
Per a 3 (8) *
Eur m 1 precursor,
Eur m 1, Eur m 1
0101 precursor, Eur
m 1 0101, Eur m 1
0102
Der f 1 precursor *,
Der f 1 *
Der m 1 *
LESLLK
TBA-1
LGDNSE
DYGKFY
VKNFIS
LRTVTP
LRTVTP
(1)
210, 212
(Der f 1)
30
(fragment)
36
(fragment)
source
Black tiger shrimp Penaeus
monodon
Timothy grass pollen Phleum
pratense
Perennial ryegrass pollen, Lolium
perenne
Shaggy cap (fungus) Coprinus
comatus
Peanut Arachis hypogaea
Antigenic?
(prediction)
novel
allerprotein
gen
-
Part of linear IgE
epitope?
-
Yes
-
Yes
-
-
-
-
No (2)
Birch pollen, Betula pendula
Cashew nut Anacardium
occidentale
Hazelnut Corylus avellana
Scab mite Sarcoptes scabiei type
hominis
Peanut Arachis hypogaea
Horse Equum caballum
-
-
No (4)
-
-
-
-
Mould Cladosporium herbarum
Housedust mite Dermatophagoides
farinae
Cockroach Periplaneta americana
Mite Euroglyphus maynei
-
-
-
-
(9)
Housedust mite Dermatophagoides
farinae
Housedust mite Dermatophagoides
microceras
Nematode Toxocara canis
-
-
(9)
-
-
(9)
-
-
(7)
Accessions:
Pen m 2: gi 27463265
Phl p 11: gi 23452313
Lol p 11: gi 626029, gi 1582249
Cop c 2: gi 11135375, gi 5689669
Ara h 2: gi 15418705
Ara h 2.02: gi 26245447
Bet v 3: gi 488605, gi 1076247, gi 1168696
Ana o 1: gi 21914823, gi 21666498
Cor a 11: gi 19338630
major allergen 1 (Sarcoptes scabiei): gi 27462848
Ara h 4: gi 5712199
Equ c 1: gi 3121758, gi 1575778
Equ c 1 chain A: gi 7766863
Cla h 5: gi 467629, gi 1086046, gi 1168970
Der f 16: gi 21591547
Per a 3: gi 1531589
Eur m 1 precursor: gi 14424447
Eur m 1: gi 102835, gi 4377538
Eur m 1 0101 precursor: gi 3941388
Eur m 1 0101: gi 3941392
Eur m 1 0102: gi 3941390
Der f 1 precursor: gi 627141, gi 730035, gi 27530349,
Der f 1: gi 7413, gi 84698, gi 9837301
Der m 1: gi 84699, gi 127205
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Annex Kleter & Peijnenburg
Potential linear IgE-epitopes in novel proteins
(2)
(3)
(4)
(5)
(6)
(7)
(8)
(9)
TBA-1: gi 299551, gi 477888
Peptide fragment (15 aa length) of Ara h 2 containing QSDRLQ sequence was not recognised by antisera pool from
15 peanut-allergic patients [Stanley et al., 1997]
Denoted by accession file as vicilin-like protein
DENEKL sequence was not in the 15 aa overlapping peptides of the Ana o 1 sequence that reacted with 3 pools of
antisera from a total of 12 cashew-allergic patients [Wang et al., 2002]
Denoted by accession file as 48-kDa glycoprotein precursor
Ara h 4 denoted by accession file as glycinin
Corresponding sequence VHRFDEG of the related allergen Ara h 3 did not bind IgE when presented as part of
overlapping 15 aa peptides of the Ara h 3 sequence to a pool of antisera from allergic patients [Rabjohn et al., 1999]
Per a 3 denoted by accession file as Cr-PI
Corresponding sequence MRTVTP in related allergen Der p 1 was part of a peptide that bound IgE and that was
considered to be part of a discontinuous epitope [Greene and Thomas, 1992]
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Annex Kleter & Peijnenburg
Potential linear IgE-epitopes in novel proteins
16. dsRNA, Vicia faba (fava bean)
Length:
Accession:
Reference amino acid sequence:
RNA editing:
Results:
Identical sequence
5825 aa
gi 7460238
Pfeiffer [1998]
not applicable
20 identical sequences, see table
Allergen (1)
name
KKLTAT
KYESNK
KYESNK
Tropomyosin
Eur m 14 (3) *
Der p 14 *
length
aa
284
1668
1662
KYESNK
Der f 14 (4) *
349
LSLVTK
LSLVTK
Eur m 14 (3) *
Der p 14 *
1668
1662
LSLVTK
Mag
341
LEKATA
LVTKEL
Blo t 11 *
Glyoxalase I
875
291
QRVVFI
GARVLVL (5)
EQSKAE
IIKSTS
Can f 1 *
Fel d 1 *
Hol l 5
Pru av 1 *, Pru av 1
chain A
Der f 3 precursor *,
Der f 3 *
Gal d 2 *, Gal d 2
chain A mutant, Gal
d 2 chain B mutant
Cor a 11 (7) *
Tyr p 2 precursor *
174
92
240
160
(Pru av 1)
232
(Der f 3)
386
(Gal d 2)
LQRVDI
SEKMKI
LSSSSG
EVKVTG
448
141
Antigenic?
(prediction)
novel
protein
(2)
-
allergen
-
-
-
-
-
-
-
-
-
-
- (6)
-
Housedust mite Dermatophagoides
farinae
Chicken Gallus domesticus
-
-
-
-
Hazelnut Corylus avellana
Storage mite Tyrophagus
putrescentiae
Mould Aspergillus fumigatus
Lobster Panulirus stimpsoni
Shrimp Metapenaeus ensis
-
-
(8)
-
Yes (9)
Yes (9)
source
Scallop Chlamys nobilis
Mite Euroglyphus maynei
European housedust mite
Dermatophagoides pteronyssinus
American housedust mite
Dermatophagoides farinae
Mite Euroglyphus maynei
European housedust mite
Dermatophagoides pteronyssinus
American housedust mite
Dermatophagoides farinae
Mite Blomia tropicalis
Rice seed, Oryza sativa (japonica
cultivar-group)
Dog Canis familiaris
Cat Felis domesticus
Velvet grass pollen Holcus lanatus
Sweet cherry Prunus avium
SSSSGV
Asp f 4 *
286
ESLLKAN
Pan s 1
274
ESLLKAN
Met e 1 *
274
(1) Accessions:
Tropomyosin: gi 9954253
Eur m 14: gi 6492307
Der p 14: gi 20385544
Der f 14: gi 1545803
Mag: gi 487661, gi 729979
Blo t 11: gi 21954740
Glyoxalase I: gi 4126809, gi 16580747
Can f 1: gi 2598974, gi 3121745
Fel d 1: gi 163825, gi 1082944
Hol l 5: gi 2266623
Pru av 1: gi 1513216, gi 7388028
Pru av 1 chain A: gi 13787043
Der f 3 precursor: gi 1311457, gi 2133564, gi 2507248
Der f 3: gi 1314736
Gal d 2: gi 129293
Gal d 2 chain A mutant: gi 15826578
Gal d 2 chain B mutant: gi 15826579
Cor a 11: gi 19338630
Tyr p 2: gi 2182106, gi 3182907
Asp f 4: gi 18202095
Pan s 1: gi 3080761, gi 14285797
Met e 1: gi 607633, gi 6094504
(2) No calculation possible for part of KKLTAT sequence at N-terminal end of novel protein
(3) Denoted by accession file as M-177 precursor
(4) Denoted by accession file as Mag3
(5) Sequence GARVLVL part of signal peptide of Fel d 1
(6) C-terminal side of sequence EQSKAE immediately flanks highest point in antigenicity plot of Hol l 5
(7) Denoted by accession file as 48-kDa glycoprotein precursor
Part of linear IgE
epitope?
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Annex Kleter & Peijnenburg
Potential linear IgE-epitopes in novel proteins
(8)
(9)
Analogous allergen Lep d 2 looses IgE-binding capability after impairment of disulfide bridging, indicative of non-linear IgEbinding epitopes [Olsson et al., 2001]. In addition, 15 aa-peptides of the Lep d 2 sequence did not bind IgE from antisera of
five mite-allergic patients [Elfman et al., 1998].
15-mer peptide from shrimp allergen Pen a 1 sequence containing ESLLKAN was bound by one out of 18 shrimp-allergic
patients’ antisera, while another 15-mer peptide containing ESLLKAN was not recognised by any antisera [Ayuso et al.,
2002]
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Annex Kleter & Peijnenburg
Potential linear IgE-epitopes in novel proteins
Identical stretches not included as positive results
Sequence
Novel protein
Protein bearing identical stretch
name
immunoglobulin gamma chain
(BAB5-L, human)
T-cell receptor alpha chain Vregion (V-J-C, human)
FLLLLW
ORFA501 (chives)
NAVDLL
ORFB (carrot)
LAGELV
ORFH522 (sunflower)
VPSGGS
FCIKFP
ORF239 (common
bean)
ORF239 (common
bean)
ORF256 (wheat)
QLSELA
ORF13 (maize)
probable polyprotein allergen
Hgg-14 (Heterodera glycines)
KILVNE
dsRNA (rice)
INLELY
dsRNA (rice)
DLNGNA
dsRNA (rice)
DLNGNA
dsRNA (rice)
LLLLTS
dsRNA (rice)
LLLLTS
dsRNA (rice)
allergen-like protein BRSn20
(Arabidopsis thaliana)
ferritin heavy chain-like protein
(Dermatophagoides
pteronyssinus)
Similar to latex allergen from
Hevea brasiliensis
(Arabidopsis thaliana)
Similar to latex allergen from
Hevea brasiliensis
(Arabidopsis thaliana)
major allergen 1-like; lacrimal
gland protein (Mus musculus)
similar to major allergen 1-like;
lacrimal gland protein (Mus
musculus)
CCGHYV
dsRNA (fava bean)
SITDIV
dsRNA (fava bean)
RPDREK
dsRNA (fava bean)
RARQSI
dsRNA (fava bean)
QGTMVT
dsRNA (fava bean)
ILSKEK
dsRNA (fava bean)
LGGLNT
dsRNA (sand pear)
probable polyprotein allergen
Hgg-14 (Heterodera glycines)
DVLSET
dsRNA (sand pear)
Llpr10.1a (Lupinus luteus)
LRVLKA
similar to latex allergen from
Hevea brasiliensis
(Arabidopsis thaliana)
60s Acidic ribosomal protein
(Caenorhabditis elegans)
Peroxiredoxin (24.9 kD)
(Caenorhabditis elegans)
Niemann-Pick disease, type
C2 (Danio rerio)
SCP-like extracellular protein,
LONg body length LON-1
(Caenorhabditis elegans)
parvalbumin beta protein
(Rana sp. CH-2001; Rana
esculenta)
similar to major allergen 1-like;
lacrimal gland protein (Mus
musculus)
immunoglobulin kappa light
chain (human)
immunoglobulin epsilon chain
(human)
similar to latex allergen from
Hevea brasiliensis
(Arabidopsis thaliana)
Remark
accession
gi 3928186
gi 758610
gi 758615
gi 763127
gi 3402683
gi 25141400 protein belongs to family of proteins
that includes allergens
gi 17554494 protein belongs to family of proteins
that includes allergens
gi 27545197 protein shows similarities to a protein
family involved with lipid recognition
that includes allergens
gi 18677166 protein identified as probable allergen
based on amino acid sequence
similarities with nematode allergen
ABA-1 [Gao et al., 2001]
gi 21553842
gi 15072346 protein described as non-allergen
gi 21554076
gi 2739381
gi 17065136
gi 20259914
gi 10181188
gi 20934683
gi 20948936
gi 20948528
gi 20956975
gi 20973491
gi 25032002
gi 28478161
gi 17553494
gi 17553496
gi 25152554
gi 20797081
gi 20797085
protein described as similar to venom
allergens
recombinant beta parvalbumin is not
recognised by antisera of patients
allergic to frog (Rana) [Hilger et al.,
2002]
gi 20824827
gi 20387060
gi 1514575
gi 2739381
gi 17065136
gi 20259914
gi 21554076
gi 18677166 protein identified as probable allergen
based on amino acid sequence
similarities with nematode allergen
ABA-1 [Gao et al., 2001]
gi 22218768 protein belongs to family of proteins
that includes allergens [Sikorski et al.,
1999]
12
Annex Kleter & Peijnenburg
Potential linear IgE-epitopes in novel proteins
References to the Annex
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Potential linear IgE-epitopes in novel proteins
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Annex Kleter & Peijnenburg
Potential linear IgE-epitopes in novel proteins
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