saporin

Chen LQ, Lv XJ, Guo QH, Lv SS, Lv N, Xu WD, Yu J, Zhang YQ (2023) Asymmetric activation of microglia in the hippocampus drives anxiodepressive consequences of trigeminal neuralgia in rodents. Br J Pharmacol 180(8):1090-1113. doi: 10.1111/bph.15994 PMID: 36443951

Objective: To determine whether and how microglia are involved in trigeminal neuralgia-induced anxiodepression.

Summary: Findings suggest that priming of microglia with ATP/P2X7 receptors in the ipsilateral hippocampus drives pain-related anxiodepressive-like behaviors via IL-1β. An asymmetric role of the bilateral hippocampus in trigeminal neuralgia-induced anxiety and depression was uncovered.

Usage: Mac-1–SAP (IT-06, 2.5 μg/1 μl, per side) was bilaterally injected into the hippocampal CA1 area on Day 0 and Day 7 after constriction of the infraorbital nerve, respectively. Unconjugated saporin was used as a control.

Related Products: Saporin (Cat. #PR-01), Mac-1-SAP mouse/human (Cat. #IT-06)

Singh SA, Vellapandian C (2023) The promising guide to LC–MS analysis and cholinesterase activity of Luffa cylindrica (L.) fruit using in vitro and in-silico analyses. Futur J Pharm Sci 9:33. doi: 10.1186/s43094-023-00478-0

Objective: Identify and analyze the extract of the plant Luffa cylindrica for bioactive and biochemical properties, particularly as it relates to bioactivity in neurological diseases.

Summary: Luffa cylindrica contains a total of 80 compounds that were identified in the ethanolic extract from LC–MS analysis. The bioactive compounds were screened for activity in receptors responsible for causing oxidative stress-associated Alzheimer’s disease. Perlolyrine was chosen to perform in-silico docking. An in vitro activity of cholinesterase showed highest inhibition at 500 μg/ml. In-silico docking of perlolyrine showed better binding affinity and score. Results revealed that out of 10 docked receptors, amyloid beta showed the highest binding affinity with an energy of −46.1 kcal/mol showing promising drug for Alzheimer’s disease. The study reports the presence of a promising, bioactive compound (perlolyrine) with promisng applications in vivo, oxidative stress-related Alzheimer’s disease.

Related Products: Saporin (Cat. #PR-01)

See Also:

• Santosh et al (2015) Mechanism of Anti-HIV Activity of Ribosome Inactivating Protein, Saporin. Protein & Peptide Letters. 22(6): 497-503. doi: 10.2174/0929866522666150428120701

Son H, Shin J, Park J (2023) Recent progress in nanomedicine-mediated cytosolic delivery. RSC Adv 13(15):9788-9799. doi: 10.1039/D2RA07111H PMID: 36998521

Summary: Cytosolic delivery of drugs, like saporin, through nanocapsules offers ways to move proteins through the body with greater specificity and efficacy. A nanoparticle that released a modified saporin protein at pH conditions correlative conditions to a tumor micro-environment was discussed as a model to deliver peptide-drugs to the cytosol.

Mohammad Hadi L, Stamati K, Yaghini E, MacRobert AJ, Loizidou M (2023) Treatment of 3D in vitro tumoroids of ovarian cancer using photochemical internalisation as a drug delivery method. Biomedicines 11(2):572. doi: 10.3390/biomedicines11020572 PMID: 36831108

Objective: To examine the efficacy of Photochemical internalization (PCI) using a porphyrin photosensitiser and a cytotoxin (Saporin) on ovarian cancer tumouroids, with HEY ovarian cancer cells in the central cancer compartment, and HDF fibroblast cells and HUVEC endothelial cells in the surrounding stromal compartment.

Summary: The efficacy was compared to tumouroids treated with either Saporin or PDT alone, or no treatment. PCI treatment was shown to be effective in the tumouroids (determined through viability assays and imaging) and caused a considerable decrease in the viability of cancer cells both within the central cancer mass and those which had migrated into the stroma, as well as a reduction in the cell density of surrounding HUVEC and HDF.

Usage: Saporin (PR-01, 20 nM) was carried out on complex tumouroids.

Related Products: Saporin (Cat. #PR-01)

Hankerd K, Koo H, McDonough KE, Wang J, Pariyar R, Tang SJ, Chung JM, La JH (2023) Gonadal hormone-dependent nociceptor sensitization maintains nociplastic pain state in female mice. Pain 164(2):402-412. doi: 10.1097/j.pain.0000000000002715 PMID: 35975896

Objective: To determine whether gonadal hormones are necessary for the development and maintenance of a peripherally maintained nociplastic pain state in female mice.

Summary: Ovariectomized females still developed a nociplastic pain state, but it was not maintained by peripheral afferent activity or spinal microglia. The findings indicate that gonadal hormones are essential for sustaining this pain state by promoting persistent sensitization of AITC-responsive afferents.

Usage: Mac-1-SAP (IT-06) or unconjugated saporin (PR-01) was administered intrathecally at 8.85 µM in 5 µL to evaluate the role of spinal microglia in nociplastic pain maintenance.

Related Products: Mac-1-SAP mouse/human (Cat. #IT-06), Saporin (Cat. #PR-01)

Q: I read on your website that, “There are two types of RIPs: type I, which are much less cytotoxic due to the lack of the B chain and type II, which are distinguished from type I RIPs by the presence of the B chain and their ability to enter cells on their own.”

In the IT-27 Streptavidin-ZAP product, which type of saporin is there? Is it both type I and type II because the saporin is purified from the plant, or is it one specific type only in the product.

A: All saporin molecules are Type I ribosome-inactivating proteins. We only use saporin. An example of a Type II RIP is ricin, which can enter a cell on its own and has been used throughout history as a method of assassination.

Streptavidin-ZAP is streptavidin attached to saporin. On its own it has no way to get inside a cell. By mixing Streptavidin-ZAP with a biotinylated molecule that is recognized on the cell surface, the resulting conjugate is able to bind and internalize saporin into a cell. Once inside saporin inactivates the ribosomes which causes cell death.

Yamaguchi S, Yamamoto K, Yamamoto R, Takamori S, Ishiwatari A, Minamihata K, Nagamune T, Okamoto A (2022) Intracellular protein photoactivation using sterically bulky caging. Chembiochem 23(22):e202200476. doi: 10.1002/cbic.202200476 PMID: 36173993

Objective: To develop an intracellular protein photoactivation method using sterically bulky caging.

Summary: Saporin was biotinylated and conjugated to streptavidin to block the active site of saporin. This temporarily inactivated protein was then activated via the cleaving of the streptavidin linker through light. This simple and versatile photoactivation method is a promising tool for studying spatio-temporal cellular events.

Usage: A sterically bulky caged Sap (cSap) was prepared via the two-step caging method using a biotinylated caging reagent (BCR). In the first step, Saporin (PR-01) was randomly modified with BCR via the amide coupling reaction. In the second step, streptavidin was conjugated with the biotin moiety on the BCR-modified Sap.

Related Products: Saporin (Cat. #PR-01)

Dzhumashev D, Timpanaro A, Ali S, De Micheli AJ, Mamchaoui K, Cascone I, Rössler J, Bernasconi M (2022) Quantum dot-based screening identifies F3 peptide and reveals cell surface nucleolin as a therapeutic target for rhabdomyosarcoma. Cancers (Basel) 14(20):5048. doi: 10.3390/cancers14205048

Objective: To investigate targeted treatment options to improve overall survival rates and to limit long-term side effects of Rhabdomyosarcoma (RMS).

Summary: The authors tested 20 different tumor-targeting peptides for their ability to bind to two RMS cell lines, Rh30 and RD, using quantum dots Streptavidin and biotin-peptides conjugates as a model for nanoparticles. F3 peptide showed the strongest binding to all RMS cell lines tested, low binding to normal control myoblasts and fibroblasts, and efficient internalization into RMS cells demonstrated by the cytoplasmic delivery of the Saporin toxin. Results indicate that nucleolin-targeting by F3 peptide represents a potential therapeutic approach for RMS.

Usage: ZAP-conjugates were prepared for selected peptides. For conjugation, Streptavidin ZAP (IT-27) was incubated with biotinylated peptides at a ratio of 1:4. After 30 min of incubation at room temperature (25°C) with gentle agitation, ZAP-conjugates were added to 2500 fast-growing cells (RD) or 5’000 slower growing (Rh30, Rh4, TTC-442) cells seeded in a 96-well plate. Three-fold dilution of initial stock was performed to obtain the series of decreasing concentrations of ZAP-conjugates (54, 18, 6, 2, 0.67, 0.22, 0.074, 0.0247 nM). ZAP-conjugates were incubated for 48 h. Viability was measured using the MTT assay.

Related Products: Streptavidin-ZAP (Cat. #IT-27), Saporin (Cat. #PR-01)

Hauf S, Rotrattanadumrong R, Yokobayashi Y (2022) Analysis of the sequence preference of saporin by deep sequencing. ACS Chem Biol 17(9):2619-2630. doi: 10.1021/acschembio.3c00733 PMID: 35969718

Objective: To study sequence-specific depurination of oligo nucleotides caused by saporin.

Summary: Data shows the sequence preference of saporin for different substrates and show that the GAGA motif is not efficiently targeted by this protein, neither in RNA nor in DNA. Instead, a preference of saporin for certain hairpin DNAs was observed.

Usage: Depurination activity of Saporin on DNA and RNA substrates (500 fmol [17nM] of saporin, while 5 pmol [167nM] of saporin was required for the RNA substrate). Nonspecific DNA:Adenosine Glycosidase Activity (150nM saporin)

Related Products: Saporin (Cat. #PR-01)

Ardini M, Vago R, Fabbrini MS, Ippoliti R (2022) From immunotoxins to suicide toxin delivery approaches: Is there a clinical opportunity?. Toxins (Basel) 14(9):579. doi: 10.3390/toxins14090579 PMID: 36136517

Objective: To give an overview describing some of the bacterial and plant enzymes studied so far for their delivery and controlled expression in tumor models.

Summary: “Suicide gene” therapy (SGT), consists of the selective delivery of genes coding for toxic proteins, into target cancer cells. This new and promising approach may overcome some of the issues related to the use of chemical agents (chemotherapy) such as as specificity, high dosages with accompanying side effects, and chemoresistance induction.