| Select | PubMed ID | Description |
| 54382 | J Allergy Clin Immunol. 1976 Jan;57(1):29-40.
Allergy to insect stings. II. Phospholipase A: the major allergen in honeybee
venom.
Sobotka AK, Franklin RM, Adkinson NF Jr, Valentine M, Baer H, Lichtenstein LM.
In order to determine the proteins of major allergenic importance in honeybee
venom (Apis mellifera) it was chromatographed on G-50 Sephadex. The four major
protein peaks eluted were identified as hyaluronidase, phospholipase, melittin,
and apamin. Testing these preparations on the leukocytes of 6 honeybee-sensitive
patients, with the in vitro method of histamine release, revealed that all
individuals were most sensitive to phospholipase A. IgE antibodies against
phospholipase A (RAST) were found in the sera of honeybee-sensitive patients and
IgG antibodies to this venom component were found in the sera from beekeepers and
venom-treated patients. Melittin appeared to be allergenic in several patients,
but the results were variable and were possibly due to contamination with
phospholipase. All patients were insensitive to the hyaluronidase and apamin
preparations. We conclude that phospholipase A is the major allergen of honeybee
venom and, since this protein is readily available, it should be useful for
diagnostic and therapeutic studies as well as for the standardization of
materials used in the management of honeybee-sensitive patients.
PMID: 54382 [Indexed for MEDLINE]
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| 6698011 |
Eur J Biochem. 1984 Mar 1;139(2):217-23.
The purification and characterisation of hyaluronidase from the venom of the
honey bee, Apis mellifera.
Kemeny DM, Dalton N, Lawrence AJ, Pearce FL, Vernon CA.
Hyaluronidase has been purified from the venom of the honey bee, Apis mellifera.
The purification proved remarkably difficult, requiring a large number of
chromatographic steps culminating in the removal of traces of phospholipase A2
with an affinity purified rabbit anti-phospholipase A2 immunosorbent column. The
purified enzyme showed a 1143-fold increase in specific activity and was
homogeneous. Electrophoresis in polyacrylamide gels (12%) containing sodium
dodecyl sulphate (pH 8.9) or urea (pH 2.8) and electrofocusing in polyacrylamide
(5%) gave a single band. The final product contained less than 0.1% phospholipase
A2 and less than 1.5% acid phosphatase and gave a single line of precipitation
against rabbit anti-hyaluronidase but was not precipitated by rabbit
anti-phospholipase A2. Previous reports of instability were not confirmed, and we
found the enzyme to be highly stable over a wide range of temperature and pH, and
to denaturing agents. Purified hyaluronidase was found to be 'sticky' when highly
pure and at low concentration, and adhered strongly to Sephadex G-75. The
relative molecular mass was estimated at 35 000-37 000 by gel filtration, and at
41 000 by sodium dodecyl sulphate/polyacrylamide gel electrophoresis. A value of
50 000 was obtained by ultracentrifugation assuming a partial specific volume of
73 cm3/g. Hyaluronidase was found to be a minor allergen in bee venom allergic
patients.
PMID: 6698011 [Indexed for MEDLINE]
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| 17479607 |
Allergy Asthma Proc. 2007 Mar-Apr;28(2):210-5.
Characterization of the N-glycans of recombinant bee venom hyaluronidase (Api m
2) expressed in insect cells.
Soldatova LN(1), Tsai C, Dobrovolskaia E, Marković-Housley Z, Slater JE.
We assessed both the monosaccharide and the oligosaccharide content of
recombinant Api m 2 using fluorophore-assisted carbohydrate electrophoresis and
HPLC. To identify the amino acid residues at which glycosylation occurs, we
digested recombinant Api m 2 with endoproteinase Glu-C and identified the
fragments that contained carbohydrate by specific staining. Recombinant Api m 2
expressed in insect cells contains N-acetylglucosamine, mannose, and fucose, as
well as trace amounts of glucose and galactose, and the oligosaccharide analysis
is consistent with heterogeneous oligosaccharide chains consisting of two to
seven monosaccharides. No sialic acid or N-acetylgalactosamine were detected.
These results are similar to published data for native Api m 2, although some
monosaccharide components appear to be absent in the recombinant protein.
Analysis of proteolytic digests indicates that of the four candidate
N-glycosylation sites, carbohydrate chains are attached at asparagines 115 and
Recombinant Api m 2 expressed in insect cells has enzymic activity and IgE
binding comparable with the native protein, and its carbohydrate composition is
very similar.
PMID: 17479607 [Indexed for MEDLINE]
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