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1TEJ from Echis Carinatus: A Heterodimeric Disintegrin Protein

Kit Ross '24 and Zion Snardon '23


Contents:


I. Introduction

As a disintegrin, 1TEJ is a transmembrane protein that binds to and antagonizes cell-surface proteins known as integrins. Integrins are important for blood clotting, which is why the disintegrin-containing venom of E. carinatus is deadly. However, integrins are also involved in promoting the invasion of tumors, giving disintegrins interesting potential applications as therapeutics. A conserved region among disintegrins called the Arg-Gly-Asp loop is known to be the site where the protein interacts with integrins. 1TEJ contains two of these loops, one in each chain, and is thereby uniquely able to inhibit two integrin proteins at once.


II. General Structure

1TEJ is a heterodimer composed of each containing 64 amino acids. The total protein is therefore 128 amino acids long and it has a molecular weight of 14.08 kDa. The heterodimeric structure of 1TEJ is composed of two very similar but distinct monomers. Because the 1TEJ monomers are attached at their , the molecule shows a distinct polarity and asymmetry. The two monomers are held together by one between Cys 12 (Chain A) & Cys 7 (Chain B) and the other between Cys 7 (Chain A) & Cys 12 (Chain B). In addition to these crossed disulfide bonds, there is also hydrogen bonding in the Asn 4 of either chain.

The A Chain contains three pairs of hydrogen bonded, antiparallel . The B Chain only contains two. Beta sheets have important structural roles in enzymes and proteins because they are able to make many hydrogen bonds to one another to form beta pleats and beta sheets. The asymmetry of Chain A and Chain B has a significant effect on its binding because Chain B is more flexible than Chain A.


III. Integrin Binding

Integrins are transmembrane proteins that process outside stimuli and communicate with the ECM and can help cell growth, adhesion, communication, etc. They are heterodimeric proteins with two distinct binding sites: the alpha (a) and beta (b). Our disintegrin has specifically high affinity when binding to aIIb3 or avb3 integrins. These two types of integrins are able to recognize conserved Arg-Gly-Asp loops in proteins that our body makes such as fibronectin and lamina, but also they recognize the Arg-Gly-Asp loops of disintegrins .


The main function of disintegrins is to bind and inhibit integrins through the action of the conserved Arg-Gly-Asp loop. 1TEJ has two of located near the C-terminus ends of each of the chains. The Arg-Gly-Asp loops on each chain are identical, except for the fact that the on the A Chain is facing the opposite direction as the one on the B Chain. The ability of this loop to bind integrins depends on how it interacts with the nearby of the chain. The proximity of these two areas of the protein brings many together that create patches of polarity that promote the binding of integrins.


IV. Integrin Specificity and Targeting

Although the Arg-Gly-Asp loop is conserved among distintegins, different disintegrins are able to target different types of integrins. Slight differences in the shape and arrangement of the Arg-Gly-Asp loop rather than sequence differences are thought to cause differing integrin specificity among disintegrin. For example, the of the Arg-Gly-Asp loop can be in an extended or narrow conformation, which enable the loop to interact with aIIb3 or avb3, respectively. Because 1TEJ is a heterodimer, the A Chain and B Chain have slightly of the Gly residue in each of their Arg-Gly-Asp loops that remarkably allows for the binding of both types of integrins.


V. References

D A Warrell, Davidson NMcD, B M Greenwood, L D Ormerod, H M Pope, B J Watkins, C R Prentice. Poisoning by bites of the saw-scaled or carpet viper (Echis carinatus) in Nigeria. National Library of Medicine. 1977 Jan; 46(181):33-62.

Iain D. Campbell and Martin J. Humphries. Integrin Structure, Activation, and Interactions. Cold Spring Harb Perspect Biol. 2011 Mar; 3(3): a004994. doi: 10.1101/cshperspect.a004994.

Gunasekera, James S Nowick. Exploring beta-sheet structure and interactions with chemical model systems. National Library of Medicine. 2008 Oct; 41(10):1319-30. doi: 10.1021/ar800064f. Epub 2008 Sep 18.

Sameeta Bilgrami, Savita Yadav, Punit Kaur, Sujata Sharma, Markus Perbandt, Christian Betzel, and Tej P. Singh. Crystal Structure of the Disintegrin Heterodimer from Saw-Scaled Viper (Echis carinatus) at 1.9 A Resolution. Biochemistry. 2005; 44 (33): 11058-11066. doi: 10.1021/bi050849y.

Sameeta Bilgrami, Shailly Tomar, Savita Yadav, Punit Kaur, Janesh Kumar, Talat Jabeen, Sujata Sharma and Tej P. Singh. Crystal Structure of Schistatin, a Disintegrin Homodimer from Saw-scaled Viper (Echis carinatus) at 2.5 A Resolution. Journal of Molecular Biology. 2004; 341(3): 829-837. doi: 10.1016/j.jmb.2004.06.048.

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