International Journal of Drug Discovery, ISSN: 0975–4423, Volume 1, Issue 1, 2009, pp-14-17 Copyright © 2009, Bioinfo Publications, International Journal of Drug Discovery, ISSN: 0975–4423, Volume 1, Issue 1, 2009 Immunoproteomics approach for prediction of antigenic epitope of Tertiapin from Apis mellifera Gomase V. S., Phadnis A. C. and Ghatak A.A. Department of Bioinformatics, Padmashree Dr. D. Y. Patil University, Plot No-50, Sector-15, CBD Belapur, Navi Mumbai, 400614, India, Mail- virusgene1@yahoo.co.in, Mobile- +91-9987770696, www.bioinfo.in Abstract –The bee venom is used for treating a wide variety of disease conditions. It has also been found to be a strong immunological agent, stimulating the body's protective mechanisms against disease. The major treatment is gene therapy or recombinant DNA vaccines involved targeting multiple antigenic components to direct and empower the immune system to protect the host from infection. Limitation of therapy to the treatment of patients suffering from various adverse reaction and contraindications are always experienced. Tertiapin is a neurotoxin from the honeybee venom. It interacts specifically with calmodulin in the presence of Ca2+. Antigenic epitopes on Tertiapin protein of Apis mellifera (honey bee) is important determinants sites for protection against disorders. As our knowledge of the immune responses to a protein antigen progressed, it became clear that the whole protein is not necessary for raising the immune response, but small segments (NCNRIIIPHMCWK, 4-16) of tertiapin protein called the antigenic determinants or the epitopes are sufficient for eliciting the desired immune response. Immunization cassettes should be capable of immunizing of broad immunity against both humoral and cellular epitope thus giving vaccines the maximum ability to deal with Tertiapin protein of Apis mellifera immune escape. We have predicted a successful immunization strategy. Keywords- Tertiapin, apitherapy, Antigenic epitope, Hydrophobicity, Hydrophilicity Introduction Apitherapy, or bee therapy, is the use of products of the common honeybee for therapeutic purposes. Honeybee venom, bee pollen, raw honey, royal jelly, and propolis are the products generally considered to have medicinal effects. These products are said to be effective against a wide range of ailments, from arthritis and chronic pain to multiple sclerosis and cancer, although few scientific studies have as yet proved their benefits. Bee venom is a potent anti-inflammatory antioxidant, antifungal, antibacterial, and possesses radioprotectant actions. It has been found to exert powerful actions as an antibacterial agent, anti-inflammatory, antiarthritic, antirheumatic, in neurodegenerative disease, as a cardiotonic, an antioxidant, and as a diaphoretic and diuretic. It has also been found to be a strong immunological agent, stimulating the body's protective mechanisms against disease. The Eclectic Botanical physicians considered it to be a potent alterative. Tertiapin Tertiapin is a neurotoxin from the honeybee venom. It interacts specifically with calmodulin in the presence of Ca2+. Miroshnikov, et al. 1983 was studied using calmodulin-cAMP phosphodiesterase system. Tertiapin does not affect the unstimulated basal activity of phosphodiesterase. However, it totally inhibits the enzyme-activating capacity of calmodulin. Analysis of the dose-dependent activation of phosphodiesterase by calmodulin in the presence of tertiapin indicated that inhibition is caused by the interaction of two tertiapin molecules with calmodulin (Kd 2 microM). The data obtained suggest that the toxic effect of tertiapin in nervous tissue is mediated by blockade of calmodulin function [1]. The cells contain phosphodiesterase of two types. One of them possesses a high affinity for cyclic AMP. The cell regulation is phosphodiesterase with low affinity for the substrate and its activity is modulated by the calmodulin-Ca2+ complex, cyclic GMP and NAD+ at micromolar concentrations. The effect on the phosphodiesterase system of a "quick" messenger, Ca2+ and "slow" messengers, cyclic GMP and NAD+, has the same consequences. The functional significance of such an activity modulation may be the necessity to maintain the conditions for phosphodiesterase functioning when Km much greater than cyclic AMP and the reaction rate are directly proportional to the substrate concentration. Studies of PC-12 cells revealed a chronic effect of phosphodiesterase activity change under the action of staphylococcal enterotoxin A. This protein induces differentiation of PC-12 cells and possesses a NAD+-glycohydrolase activity. As a result, the enzyme activity falls, cyclic AMP level increases and cell differentiation starts. The activity of soluble phosphodiesterase of PC-12 cells also decreases under the effect of two neurotoxins from bee venom, melittin and tertiapin. Both the toxins at concentration of 10 microM completely block calcium regulation of the enzyme. Inhibition of Ca2+ action is achieved as the result of the activated calmodulin molecule [2]. Tertiapin is a small protein derived from honeybee venom. It inhibits the GIRK1/4 and ROMK1 channels with nanomolar affinities. Methionine residue 13 in TPN interacts with residue F148 in the channel, located just outside of the narrow region of the ROMK1 pore [3]. A recent study indicated that tertiapin is a potent blocker of certain types of inwardly rectifying K (+) (Kir) channels. Tertiapin fully inhibited acetylcholine (1 microM)-induced muscarinic K (+) (K(ACh)) channel currents in atrial myocytes. At the single-channel level, tertiapin inhibited the