In vitro refolding and functional analysis of polyhistidine-tagged Buthus martensii Karsch antitumor-analgesic peptide produced in Escherichia coli

Cao, Qingxin; Lü, Wei; Cai, Xueting; Hu, Chunping; Wang, Chen; Ye, Juan; Yan, Huaijiang; Yang, Yang; Wang, Zhigang; Huo, Jiege; Liu, Yan; Yu, Ye; Ling, Changquan; Cao, Peng

doi:10.1007/s10529-015-1936-8

Cited by 6 publications

(2 citation statements)

References 14 publications

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“…This procedure allows a determined folding of the recombinant peptide resulting an active calcin. Using this system avoids the production of several isoforms as discussed before (Estrada et al, 2007; Saucedo et al, 2012; Cao et al, 2015). The use of thioredoxin as fusion protein certainly favored the folding of the disulfide bonds because it displays chaperone activity, which provides high stability to the protein with which is fused (LaVallie et al, 1993; Kern et al, 2003).…”

Section: Resultsmentioning

confidence: 99%

Recombinant expression of Intrepicalcin from the scorpion Vaejovis intrepidus and its effect on skeletal ryanodine receptors

Vargas-Jaimes

Liang

Zhang

et al. 2017

Biochimica et Biophysica Acta (BBA) - General Subjects

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Background Scorpion venoms contain toxins that modulate ionic channels, among which are the calcins, a small group of short, basic peptides with an Inhibitor Cystine Knot (ICK) motif that target calcium release channels/ryanodine receptors (RyRs) with high affinity and selectivity. Here we describe the heterologous expression of Intrepicalcin, identified by transcriptomic analysis of venomous glands from Vaejovis intrepidus. Methods Recombinant Intrepicalcin was obtained in Escherichia coli BL21-DE3 (periplasm) by fusing the intrepicalcin gene to sequences coding for signal-peptide, thioredoxin, His-tag and enterokinase cleavage site. Results [3H]Ryanodine binding, used as a functional index of RyR activity, revealed that recombinant Intrepicalcin activates skeletal RyR (RyR1) dose-dependently with Kd = 17.4 ± 4.0 nM. Intrepicalcin significantly augments the bell-shaped [Ca2+]-[3H]ryanodine binding curve at all [Ca2+] ranges, as is characteristic of the calcins. In single channel recordings, Intrepicalcin induces the appearance of a subconductance state in RyR1 with a fractional value ~55% of the full conductance state, very close to that of Vejocalcin. Furthermore, Intrepicalcin stimulates Ca2+ release at an initial dose = 45.3 ± 2.5 nM, and depletes ~50% of Ca2+ load from skeletal sarcoplasmic reticulum vesicles. Conclusions We conclude that active recombinant Intrepicalcin was successfully obtained without the need of manual oxidation, enabling it to target RyR1s with high affinity. General Significance This is the first calcin heterologously expressed in the periplasma of Escherichia coli BL21-DE3, shown to be pharmacologically effective, thus paving the way for the generation of Intrepicalcin variants that are required for structure-function relationship studies of calcins and RyRs.

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Section: Resultsmentioning

confidence: 99%

Recombinant expression of Intrepicalcin from the scorpion Vaejovis intrepidus and its effect on skeletal ryanodine receptors

Vargas-Jaimes

Liang

Zhang

et al. 2017

Biochimica et Biophysica Acta (BBA) - General Subjects

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“…Different toxins from the scorpion Buthus martensii are promising for the treatment of pain in a xenograft tumor mouse model [ 77 , 78 ]. These toxins can be important candidates also to use in acute thermal pain model induced by a hot plate, in acute inflammation model induced by formalin [ 79 , 80 , 81 , 82 ], in a mouse-twisting pain model, mechanical allodynia, in a nociception model induced by a shutter-controlled lamp, and in an inflammation model induced by formalin and acetic acid writhing [ 81 ].…”

Section: Toxins Targeting Pain: Discovering Potential Analgesicsmentioning

confidence: 99%

Unveiling the Pain Relief Potential: Harnessing Analgesic Peptides from Animal Venoms

Pereira,

Cavalcante,

Angstmam

et al. 2023

Pharmaceutics

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The concept of pain encompasses a complex interplay of sensory and emotional experiences associated with actual or potential tissue damage. Accurately describing and localizing pain, whether acute or chronic, mild or severe, poses a challenge due to its diverse manifestations. Understanding the underlying origins and mechanisms of these pain variations is crucial for effective management and pharmacological interventions. Derived from a wide spectrum of species, including snakes, arthropods, mollusks, and vertebrates, animal venoms have emerged as abundant repositories of potential biomolecules exhibiting analgesic properties across a broad spectrum of pain models. This review focuses on highlighting the most promising venom-derived toxins investigated as potential prototypes for analgesic drugs. The discussion further encompasses research prospects, challenges in advancing analgesics, and the practical application of venom-derived toxins. As the field continues its evolution, tapping into the latent potential of these natural bioactive compounds holds the key to pioneering approaches in pain management and treatment. Therefore, animal toxins present countless possibilities for treating pain caused by different diseases. The development of new analgesic drugs from toxins is one of the directions that therapy must follow, and it seems to be moving forward by recommending the composition of multimodal therapy to combat pain.

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Microbial production of toxins from the scorpion venom: properties and applications

Amorim

Cordeiro

Pinheiro-Júnior

et al. 2018

Appl Microbiol Biotechnol

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Scorpion venom are composed mainly of bioactive proteins and peptides that may serve as lead compounds for the design of biotechnological tools and therapeutic drugs. However, exploring the therapeutic potential of scorpion venom components is mainly impaired by the low yield of purified toxins from milked venom. Therefore, production of toxin-derived peptides and proteins by heterologous expression is the strategy of choice for research groups and pharmaceutical industry to overcome this limitation. Recombinant expression in microorganisms is often the first choice, since bacteria and yeast systems combine high level of recombinant protein expression, fast cell growth and multiplication and simple media requirement. Herein, we present a comprehensive revision, which describes the scorpion venom components that were produced in their recombinant forms using microbial systems. In addition, we highlight the pros and cons of performing the heterologous expression of these compounds, regarding the particularities of each microorganism and how these processes can affect the application of these venom components. The most used microbial system in the heterologous expression of scorpion venom components is Escherichia coli (85%), and among all the recombinant venom components produced, 69% were neurotoxins. This review may light up future researchers in the choice of the best expression system to produce scorpion venom components of interest.

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In vitro refolding and functional analysis of polyhistidine-tagged Buthus martensii Karsch antitumor-analgesic peptide produced in Escherichia coli

Abstract: This refolding protocol optimized for C-terminal His6-tagged scorpion rAGAP is potentially applicable to similar long-chain and cysteine-rich toxins.

Cited by 6 publications

References 14 publications

Recombinant expression of Intrepicalcin from the scorpion Vaejovis intrepidus and its effect on skeletal ryanodine receptors

Recombinant expression of Intrepicalcin from the scorpion Vaejovis intrepidus and its effect on skeletal ryanodine receptors

Unveiling the Pain Relief Potential: Harnessing Analgesic Peptides from Animal Venoms

Microbial production of toxins from the scorpion venom: properties and applications

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