Ophanin is a

toxin A toxin is a naturally occurring poison produced by metabolic activities of living cells or organisms. They occur especially as proteins, often conjugated. The term was first used by organic chemist Ludwig Brieger (1849–1919), derived ...

found in the venom of the King Cobra (''

Ophiophagus hannah The king cobra (''Ophiophagus hannah'') is a species complex of snakes endemic to Asia. With an average of and a record length of , it is the world's longest venomous snake and among the heaviest. Under the genus ''Ophiophagus'', it is not p ...

''), which lives throughout

South East Asia Southeast Asia is the geographical United Nations geoscheme for Asia#South-eastern Asia, southeastern region of Asia, consisting of the regions that are situated south of China, east of the Indian subcontinent, and northwest of the Mainland Au ...

. This

belongs to the

cysteine-rich secretory protein Cysteine-rich secretory proteins, often abbreviated as CRISPs, are a group of glycoproteins. They are a subgroup of the CRISP, antigen 5 and Pr-1 (CAP) protein superfamily and also contain a domain related to the ShK toxins. They are substantially ...

(CRISP) family. Ophanin weakly blocks the contraction of

smooth muscles Smooth muscle is one of the three major types of vertebrate muscle tissue, the others being skeletal and cardiac muscle. It can also be found in invertebrates and is controlled by the autonomic nervous system. It is non- striated, so-called becau ...

elicited by high potassium-induced

depolarization In biology, depolarization or hypopolarization is a change within a cell (biology), cell, during which the cell undergoes a shift in electric charge distribution, resulting in less negative charge inside the cell compared to the outside. Depolar ...

, suggesting that it inhibits voltage-dependent calcium channels.

Etymology

The toxin was named ''ophanin'' after the snake whose venom it is derived from, the King Cobra (''

'').

Sources

Ophanin is produced in the venom glands of the King Cobra (''O. Hannah''). Although the venom has relatively low

toxicity Toxicity is the degree to which a chemical substance or a particular mixture of substances can damage an organism. Toxicity can refer to the effect on a whole organism, such as an animal, bacteria, bacterium, or plant, as well as the effect o ...

, this is compensated by the high amounts of it injected into the prey for each bite.

Chemistry

Structure

Ophanin was successfully isolated from ''O. Hannah'' venom by

gel filtration Size-exclusion chromatography, also known as molecular sieve chromatography, is a chromatographic method in which molecules in solution are separated by their shape, and in some cases size. It is usually applied to large molecules or macromolecul ...

and cation-exchange

chromatography In chemical analysis, chromatography is a laboratory technique for the Separation process, separation of a mixture into its components. The mixture is dissolved in a fluid solvent (gas or liquid) called the ''mobile phase'', which carries it ...

. Its molecular weight is 25 kDa (from positions 19 – 239), which conforms to the molecular mass predicted from its cDNA sequences.

Homology

Ophanin is a cysteine-rich secretory protein and therefore belongs to the CRISP family. These proteins possess 16 strictly conserved cysteines and contain 8

disulfide bonds In chemistry, a disulfide (or disulphide in British English) is a compound containing a functional group or the anion. The linkage is also called an SS-bond or sometimes a disulfide bridge and usually derived from two thiol groups. In in ...

. Ten of the 16 cysteine residues are clustered at the

C-terminal The C-terminus (also known as the carboxyl-terminus, carboxy-terminus, C-terminal tail, carboxy tail, C-terminal end, or COOH-terminus) is the end of an amino acid chain (protein or polypeptide), terminated by a free carboxyl group (-COOH). When t ...

end of the protein. Ophanin belongs to the “long” CRISPs subgroup, which consists of the 9 CRISPs with the longest sequences. Snake venom CRISPs belonging to different subgroups act on different biological targets, contributing in this way to the diversity of damaging effects of snake venoms.

Family

The phylogenetic tree constructed from the nucleotide sequences of all known snake venom CRISPs shows that ophanin is more closely related to the

Viperidae Vipers are snakes in the family Viperidae, found in most parts of the world, except for Antarctica, Australia, Hawaii, Madagascar, New Zealand, Ireland, and various other isolated islands. They are venomous snake, venomous and have long (relat ...

branch than the

Elapidae Elapidae (, commonly known as elapids , from , variant of "sea-fish") is a family of snakes characterized by their permanently erect fangs at the front of the mouth. Most elapids are venomous, with the exception of the genus '' Emydocephalus ...

branch even though ''O. Hannah'' belongs to the Elapidae snakes. Ophanin, along with other specific snake toxins like triflin and ablomin, is also a helothermine-related venom protein (Helveprin) which was originally isolated from the skin of the Mexican beaded lizard.

Target

Ophanin is a weak blocker of the high potassium-induced contraction of smooth muscles. Snake venom CRISP family proteins inhibit depolarization-induced smooth muscle contraction to different extents. Compared to the normal contraction of smooth muscle, ophanin is able to reduce their force of contractility to 84% ± 1%, which is less than most other CRISPs. The differences between the inhibitory activity of CRISPs may be explained through sequence comparisons that suggest a site that may be critical for inhibition of channel activity. Phe189 and Glu186 are the most likely functional residues: strong blockers of smooth muscle contraction ( ablomin, triflin, and latisemin) all have Phe189, and all blockers of smooth muscle contraction except ophanin have Glu186. The significance of this lack of the probable functional residues in ophanin has not yet been addressed. However, it is likely that the picture is more complex and other residues contribute to the inhibitory activity of CRISPs on smooth muscle contraction and some data supports this. For example, pseudecin, while also having Phe189, does not affect depolarization-induced contraction.

Mode of action

There is no direct evidence of a particular mode of action of ophanin blocking depolarization-induced contractions of the smooth muscles. However, based on the hypothesis of Yamazaki and colleagues in regards to ablomin, another snake venom toxin from the CRISP family that also blocks depolarization-induced smooth muscle contraction, we can postulate a similar mechanism might be in place for ophanin. Since ablomin only blocks contraction induced by depolarization, but not by caffeine, the effect of ablomin is likely to be caused by inhibition of voltage-gated ion channels. An activation of smooth muscle cells through caffeine activates

ryanodine Ryanodine is a poisonous diterpenoid found in the South American plant ''Ryania speciosa'' (Salicaceae). It was originally used as an insecticide. The compound has extremely high affinity to the open-form ryanodine receptor, a group of calcium ch ...

receptors of the

sarcoplasmic reticulum The sarcoplasmic reticulum (SR) is a membrane-bound structure found within muscle cells that is similar to the smooth endoplasmic reticulum in other cells. The main function of the SR is to store calcium ions (Ca2+). Calcium ion levels are kep ...

, whereas an activation through high levels of extracellular potassium depolarizes the membrane (due to the change of the

reversal potential In a biological membrane, the reversal potential is the membrane potential at which the direction of ionic current reverses. At the reversal potential, there is no net flow of ions from one side of the membrane to the other. For channels that are pe ...

for potassium towards more positive values) and would then activate voltage-gated calcium-ion channels leading to high levels of intracellular calcium ions. The intracellular calcium ion concentration correlates well with contraction force in the rat-tail artery. Thus, contraction following extracellular application of high-potassium solution depends on the influx of the extracellular calcium ions through voltage-gated calcium channels. Therefore, ablomin (and by extension ophanin) most likely targets voltage-gated calcium channels on smooth muscle.

Toxicity

The of the venom in mice is ~1.2 to 3.5 mg/kg via intravenous injection. The LD50 of ophanin is not yet known.

References

{{reflist Ion channel toxins Snake toxins