Mellitin

Mellitin is basic polypeptide from the venom of the honey bee (Apis mellifera); it contains 62 amino acids, has cytolytic properties, causes contracture of muscle, releases histamine and disrupts surface tension, probably due to lysis of cell and mitochondrial membranes.

• Mellitin Certificate of Analysis
• Mellitin MSDS
• Mellitin Analytical Data
• Mellitin References

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Mellitin characteristics

 Mellitin is natural, non-synthetic, polypeptide from European honey bee, Apis mellifera, venom; the principle hemolytic component.

 Synonyms: melittin, melliten, melittin, melitten.

Product Name	MELITTIN
Description	Natural, non-synthetic, polypeptide from European honey bee, Apis mellifera, venom; the principle hemolytic component
Amino Acid Sequence	{GLY}{ILE}{GLY}{ALA}{VAL}{LEU}{LYS}{VAL}{LEU}{THR} {THR}{GLY}{LEU}{PRO}{ALA}{LEU}{ILE}{SER}{TRP}{ILE} {LYS}{ARG}{LYS}{ARG}{GLN}{GLN}-NH2
IDNUMBER/Order Ref.	Melittin, CAS 20449-79-0
CAS Registry Number	20449-79-0
MDL Number	MFCD00076868
Molecular Formula	C131 H229 N39 O31
Molecular Weight	2846.46
APPEARANCE	Dry fluffy powder, pale-yellow color
Original Storage Container
Storage temperature	-20°C
Transportation (short-term) temperature	Ambient
SOLUBILITY	Solubility in an aqueous medium; melittin is soluble as a tetramer in aqueous salt solutions.
PURITY (HPLC)	>97%
Known Activity	26-residue polypeptide binds calmodulin in a Ca2+ dependent manner, activates phospholipase A2, inhibits protein kinase C (IC50 = 5 – 7 mM) by binding to the catalytic domain in a Mg-ATP sensitive manner. Highly sensitive substrate for chymotrypsin detection. Please refer to below references and selected article abstracts on extensive application of Melittin.

Download Mellitin MSDS in PDF format.
Download Mellitin MSDS in PDF format.

Section 1. Product Information

Product Name	MELITTIN, Melittin-T
	Natural, non-synthetic, polypeptide from European honey bee, Apis mellifera, venom; the principle hemolytic component
Amino Acid Sequence	{GLY}{ILE}{GLY}{ALA}{VAL}{LEU}{LYS}{VAL}{LEU}{THR} {THR}{GLY}{LEU}{PRO}{ALA}{LEU}{ILE}{SER}{TRP}{ILE} {LYS}{ARG}{LYS}{ARG}{GLN}{GLN}-NH2
IDNUMBER	Melittin, CAS 20449-79-0
CAS Registry Number	20449-79-0
MDL Number	MFCD00076868
Molecular Formula	C131H229N39O31
Molecular Weight	2846.46

Section 2. Hazards Identification

EMERGENCY OVERVIEW

Harmful.

Poison. May be fatal if enters bloodstream. Sensitizer. Target organ(s): Central nervous system. Heart. Do not breathe dust. Do
not use if skin is cut or scratched. Wash thoroughly after handling.

Section 3.  First Aid Measures

ORAL EXPOSURE

If swallowed, wash out mouth with water provided person is conscious. Call a physician immediately.

INHALATION EXPOSURE

If inhaled, remove to fresh air. If not breathing give artificial respiration. If breathing is difficult, give oxygen.

DERMAL EXPOSURE

In case of skin contact, flush with copious amounts of water for at least 15 minutes. Remove contaminated clothing and shoes.

Call a physician.

EYE EXPOSURE

In case of contact with eyes, flush with copious amounts of water for at least 15 minutes. Assure adequate flushing by

separating the eyelids with fingers. Call a physician.

Section 4. Accidental Release Measures

PROCEDURE TO BE FOLLOWED IN CASE OF LEAK OR SPILL

Evacuate area.

PROCEDURE(S) OF PERSONAL PRECAUTION(S)

Wear self-contained breathing apparatus, rubber boots, and heavy rubber gloves.

METHODS FOR CLEANING UP

Spilled material should be carefully wiped up or moistened with water and removed. Ventilate area and wash spill site after

material pickup is complete.

Section 5. Handling and Storage

HANDLING

User Exposure: Avoid inhalation. Do not get in eyes, on skin, on clothing. Avoid prolonged or repeated exposure. Do not use if

skin is cut or scratched. Wash thoroughly after handling.

STORAGE

Suitable: Keep tightly closed.

Store at -20°C

Section 6. Exposure Controls / PPE

ENGINEERING CONTROLS

Safety shower and eye bath. Use only in a chemical fume hood.

PERSONAL PROTECTIVE EQUIPMENT

Respiratory: Use respirators and components tested and approved under appropriate government standards such as NIOSH (US) or CEN(EU). Where risk assessment shows air-purifying respirators are appropriate use a full-face particle respirator type N100 (US) or type P3 (EN 143) respirator cartridges as a backup to engineering controls. If the respirator is the sole means of protection, use a full-face supplied air respirator.

Hand: Compatible chemical-resistant gloves.

Eye: Chemical safety goggles.

GENERAL HYGIENE MEASURES

Wash contaminated clothing before reuse.

Section 7.  Stability and Reactivity

STABILITY

Stable: Stable.

Materials to Avoid: Strong oxidizing agents.

HAZARDOUS DECOMPOSITION PRODUCTS

Hazardous Decomposition Products: Nature of decomposition products not known.

HAZARDOUS POLYMERIZATION

Hazardous Polymerization: Will not occur

Section 8. Toxicological Information

ROUTE OF EXPOSURE

Skin Contact: May cause skin irritation.

Skin Absorption: May be harmful if absorbed through the skin.

Eye Contact: May cause eye irritation.

Inhalation: May be harmful if inhaled. Material may be irritating to mucous membranes and upper respiratory tract.

Ingestion: May be harmful if swallowed.

SENSITIZATION

Sensitization: May cause allergic reaction.

TARGET ORGAN(S) OR SYSTEM(S)

Heart. Central nervous system.

SIGNS AND SYMPTOMS OF EXPOSURE

To the best of our knowledge, the chemical, physical, and toxicological properties have not been thoroughly investigated.

May be fatal if enters bloodstream.

CONDITIONS AGGRAVATED BY EXPOSURE

May be fatal if enters bloodstream.

Section 9. Disposal Considerations

APPROPRIATE METHOD OF DISPOSAL OF SUBSTANCE OR PREPARATION

Contact a licensed professional waste disposal service to dispose of this material. Observe all federal, state, and local environmental regulations.

Section 10 Transport Information

Non-Hazardous for Transport: This substance is considered to be non-hazardous for transport.

IATA

Non-Hazardous for Air Transport: Non-hazardous for air transport.

Section 11 - Other Information 

For R&D use only. Not for internal or external human consumption.

WARRANTY

THE ABOVE INFORMATION IS BELIEVED TO BE CORRECT BUT DOES NOT PURPORT TO BE ALL INCLUSIVE AND SHALL BE USED ONLY AS A GUIDE. TIMTEC SHALL NOT BE HELD LIABLE FOR ANY DAMAGE RESULTING FROM HANDLING OR FROM CONTACT WITH THE ABOVE PRODUCT.

Mellitin HPLC spectra data

Mellitin HPLC spectra PDF

Mellitin HPLC spectra PNG

Melittin Time-of-flight (TOF) mass spectra data

Mellitin Time-of-flight (TOF) mass spectra PDF

Mellitin Time-of-flight (TOF) mass spectra PNG

Mellitin UV spectra data

Mellitin UV spectra PDF

Mellitin UV spectra PNG

References

Tosteson, M.T., Tosteson D.C. The sting. Melittin forms channels in lipid bilayers. Biophysical Journal 36: 109-116 (1981)

Abstract

Melittin, a toxin of bee venom, is a cationic polypeptide composed of 26 amino acids. The six residues of the C-terminal end are polar and 19 of the 20 residues of the N-terminal end are hydrophobic. Exposure of the lecithin bilayer to melittin results in the formation of channels that are more permeable to anions that to cations. Unilateral addition of melittin produces a voltage-dependent increase in membrane conductance when the side where the polypeptide is present in made positive but not when it is made negative. At a fixed voltage, the conductance increases with the fourth power of the melittin concentration in the aqueous phase. At a fixed peptide concentration, the conductance increases approximately e-fold per 6-mV increase in the electrical potential difference across the membrane. These results suggest that four melittin monomers are needed to form a channel and, furthermore, that a minimum of four equivalent electronic charges need to be displaced by the electrical field to explain the voltage dependence of the conductance.

Hristova K, et al. Structure, location, and lipid perturbations of melittin at the membrane interface. Biochim Biophys Acta. 1990 May 7;1031(2):143-61.

Abstract

The molecular mechanisms underlying the various effects of melittin on membranes have not been completely defined and much of the evidence described indicates that different molecular mechanisms may underlie different actions of the peptide. Ideas about the formation of transbilayer aggregates of melittin under the influence of a transbilayer potential, and for bilayer structural perturbation arising from the location of the peptide helix within the head group region of the membrane have been made based on the crystal structure of the peptide, the kinetics and concentration dependence of melittins membrane actions, together with simple ideas about the conformational properties of amphipathic helical peptides and their interactions with membranes. Physical studies of the interaction of melittin with model membranes have been useful in determining the potential of the peptide to adopt different locations, orientations and association states within membranes under different conditions, but the relationship of the results obtained to the actions of melittin in cell membranes or under the influence of a membrane potential are unclear. Experimental definition of the interaction of melittin with more complex membranes, including the erythrocyte membrane or in bilayers under the influence of a transmembrane potential, will require direct study in these membranes. Experiments employing labeled melittins for ESR, NMR or fluorescence experiments are promising both for their sensitivity (ESR and fluorescence) and the ability to focus on the peptide within the background of endogenous proteins within cell membranes. The study of melittin in model membranes has been useful for the development of methodology for determination of membrane protein structures. Despite the structural complexity of integral membrane proteins, it is interesting that in some respects their study be more straightforward, lacking as they do the elusive properties of melittin (and other structurally labile membrane peptides) which limit the possibility of defining their interaction with membranes in terms of a single conformation, location, orientation and association state within the membrane.

Terwilliger, TC., Eisenberg, D. The structure of melittin. I. Structure determination and partial refinement. J. Biol. Chem., Vol. 257, Issue 11, 6010-6015, 06, 1982

Abstract

Melittin is the principal protein component of bee venom and is thought to function as a lytic agent. Despite its predominantly hydrophobic character, melittin is soluble as a tetramer in aqueous salt solutions. We report here on the determination of the crystal structure of tetrameric melittin at 2.8-A resolution by the method of multiple isomorphous replacement,followed by partial atomic refinement at 2.0-A resolution. The melittin tetramer contains a noncrystallographic 2-fold axis of symmetry in addition to a crystallographic 2-fold axis, so that the four polypeptide chains have nearly identical structures. The noncrystallographic 2-fold axis was utilized twice during the determination of the structure. The multipleisomorphous replacement electron density map was averaged over this 2-fold axis before model building and strict noncrystallographic symmetry was assumed during the initial stages of atomic refinement. The 2.8-A resolution electron density map suggests that the melittin monomer contains two alpha- helical regions separated by a non-alpha-helical segment atresidues 11 and 12. Difference maps at 2.0-A resolution tend to confirm this structure and reveal that at least six solvent molecules are bound to the melittin tetramer in the crystal. The relatively high occupancies of four of these suggest that they are ions of crystallization rather than water molecules.

DeGrado, W.F. et al. Kinetics and mechanism of hemolysis induced by melittin and by a synthetic melittin analogue. Biophys J. 1982 January; 37(1): 329–338.

Abstract

The cytotoxic peptide from honeybee venom, melittin, and a synthetic peptide analogue of it lyse human erythrocytes in a biphasic process. The kinetics of the lysis in 0.30 M sucrose, 0.01 M sodium phosphate, pH 7.30 at 4 degrees C were investigated. Our results show that melittin rapidly binds to the outer surface of the erythrocyte membrane, and the surface-bound monomers produce transient openings through which approximately 40 hemoglobin molecules can escape. Concomitantly, the melittin loses its ability to effect the process, presumably by translocation through the bilayer. The half-life for this process is 1.2 min. In a much slower process, dimers of this internalized melittin again produce transient membrane openings in a steady state. On a molar basis, the synthetic peptide analogue produces a fast process comparable to that caused by melittin, but is more efficient in the slow phase. Escape of hemoglobin and of carbonic anhydrase through the openings is diffusion controlled. These results suggest that the functional units necessary for the activity of melittin-like cytotoxic peptides are a 20 amino acid amphiphilic alpha-helix with a hydrophobic:hydrophilic ratio greater than 1 and a short segment with a high concentration of positive charges.

Boman, H.G. et al. Antibacterial and antimalarial properties of peptides that are cecropin-melittin hybrids. Dept of Microbiology, University of Stockholm, Sweden. FEBS Lett. 1989 Dec 18;259(1):103-6.

Abstract

Solid phase synthesis was used to produce 5 hybrid peptides containing sequences from the antibacterial peptide, cecropin A, and from the bee venom toxin, melittin. Four of these chimeric peptides showed good antibacterial activity against representative Gram-negative and Gram-positive bacterial species. The best hybrid, cecropin A(1-13)-melittin(1-13) was 100-fold more active than cecropin A against Staphylococcus aureus. It was also a 10-fold better antimalarial agent than cecropin B or magainin 2. Sheep red cells were lysed by melittin at low concentrations, but not by the hybrid molecules, even at 50 times higher concentrations.

Lauterwein, J. et al. Physicochemical studies of the protein-lipid interactions in melittin-containing micelles. Biochim Biophys Acta. 1979 Sep 21;556(2):244-64.

Abstract

Complexes of melittin with detergents and phospholipids have been characterized by fluorescence, circular dichroism, ultracentrifugation, quasi-elastic light scattering and 1H nuclear magnetic resonance (NMR) experiments. By ultracentrifugation and quasi-elastic light-scattering measurements it is shown that melittin forms stoichiometrically well-defined complexes with dodecylphosphocholine micelles consisting of one melittin molecule and approximately forty detergent molecules. Evidence from fluorescence, circular dichroism and 1H nuclear magnetic resonance experiments indicates that the conformation of melittin bound to micelles of various detergents or of diheptanoyl phosphatidylcholine is largely independent of the type of lipid and furthermore appears to be quite closely related to the conformation of melittin bound to phosphatidylcholine bilayers. 1H NMR is used to investigate the conformation of micelle-bound melittin in more detail and to compare certain aspects of the melittin conformation in the micelles with the spatial structures of monomeric and self-aggregated tetrameric melittin in aqueous solution. The experience gained with this system demonstrates that high resolution NMR of complexes of membrane proteins with micelles provides a viable method for conformational studies of membrane proteins.

Ghosh, A. K. et al. Modulation of Tryptophan Environment in Membrane-Bound Melittin by Negatively Charged Phospholipids: Implications in Membrane Organization and Function. Biochemistry, 36 (47), 14291 -14305, 1997. bi971933j S0006-2960(97)01933-8

Abstract

Melittin is a cationic hemolytic peptide isolated from the European honey bee, Apis mellifera. Since the association of the peptide in the membrane is linked with its physiological effects, a detailed understanding of the interaction of melittin with membranes is crucial. We have investigated the interaction of melittin with membranes of varying surface charge in the context of recent studies which show that the presence of negatively charged lipids in the membrane inhibits membrane lysis by melittin. The sole tryptophan residue in melittin has previously been shown to be critical for its hemolytic activity. The organization and dynamics of the tryptophan residue thus become important to understand the peptide activity in membranes of different charge types. Wavelength-selective fluorescence was utilized to monitor the tryptophan environment of membrane-bound melittin. Melittin exhibits a red edge excitation shift (REES) of 5 nm when bound to zwitterionic membranes while in negatively charged membranes, the magnitude of REES is reduced to 2-3 nm. Further, wavelength dependence of fluorescence polarization and near-UV circular dichroism spectra reveal characteristic differences in the tryptophan environment for melittin bound to zwitterionic and anionic membranes. These studies are supported by time-resolved fluorescence measurements of membrane-bound melittin. Tryptophan penetration depths for melittin bound to zwitterionic and anionic membranes were analyzed by the parallax method [Chattopadhyay, A., and London, E. (1987) Biochemistry 26, 39-45] utilizing differential fluorescence quenching obtained with phospholipids spin-labeled at two different depths. Our results provide further insight into molecular details of membrane lysis by melittin and the modulation of lytic activity by negatively charged lipids.

Comte, M. et al. Ca2+-dependent high-affinity complex formation between calmodulin and melittin.Biochem J. 1983 January 1; 209(1): 269–272.

Abstract

The amphiphatic polypeptide melittin migrates as an equimolar complex with bovine brain calmodulin when monitored by gel disc electrophoresis or gel filtration in the presence of Ca2+, even in 4M-urea. The complex disassociates in the presence of EDTA and urea. The affinity is of the same order as that of calmodulin for its target enzymes, and more than 1000-fold higher than that of calmodulin for basic peptide hormones or hydrophobic drugs. The activation of brain phosphodiesterase by calmodulin is inhibited by melittin. The kinetics of inhibition suggest competition between the enzyme and melittin for calmodulin. The calmodulin-melittin interaction may constitute a model for that existing between calmodulin and its target enzymes.