Proteomic analysis of the honey bee worker venom gland focusing on the mechanisms of protection against tissue damage.
Honey bee workers use venom for the defence of the colony and themselves when they are exposed to dangers and predators. It is produced by a long thin, convoluted, and bifurcated gland, and consists of several toxic proteins and peptides. The present study was undertaken in order to identify the mechanisms that protect the venom gland secretory cells against these harmful components. Samples of whole venom glands, including the interconnected reservoirs, were separated by two-dimensional gel electrophoresis and the most abundant protein spots were subjected to mass spectrometric identification using MALDI TOF/TOF-MS and LC MS/MS. This proteomic study revealed four antioxidant enzymes: CuZn superoxide dismutase (SOD1), glutathione-S-transferase sigma 1 isoform A (GSTS1), peroxiredoxin 2540 (PXR2540) and thioredoxin peroxidase 1 isoform A (TPX1). Although glutathione-S-transferase (GST) has also been associated with xenobiotic detoxification, the protein we found belongs to the GST Sigma class which is known to protect against oxidative stress only. Moreover, we could demonstrate that the GST and SOD activity of the venom gland was low and moderate, respectively, when compared to other tissues from the adult honey bee. Several proteins involved in other forms of stress were likewise found but it remains uncertain what their function is in the venom gland. In addition to major royal jelly protein 9 (MRJP9), already found in a previous proteomic study, we identified MRJP8 as second member of the MRJP protein family to be associated with the venom gland. Transcripts of both MRJPs were amplified and sequenced. Two endocuticular structural proteins were abundantly present in the 2D-gel and most probably represent a structural component of the epicuticular lining that protects the secretory cells from the toxins they produce.
Laboratory of Zoophysiology, Ghent University, K.L. Ledeganckstraat 35, B-9000 Ghent, Belgium.
Nico
N
Peiren
Dirk C
DC
de Graaf
Frank
F
Vanrobaeys
Ellen L
EL
Danneels
Bart
B
Devreese
Jozef
J
Van Beeumen
Frans J
FJ
Jacobs
D000595:Amino Acid Sequence; D000818:Animals; D001514:Bee Venoms / Q000633:toxicity*; D001516:Bees / Q000737:chemistry / Q000502:physiology*; D015180:Electrophoresis, Gel, Two-Dimensional; D005982:Glutathione Transferase / Q000032:analysis; D019476:Insect Proteins / Q000032:analysis*; D008969:Molecular Sequence Data; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013482:Superoxide Dismutase / Q000032:analysis
D001514:Bee Venoms; D019476:Insect Proteins; D013482:Superoxide Dismutase; D005982:Glutathione Transferase