Tag Archives: Peptides

Protease OMA1 Activity is Measured by MCA Fluorescent Peptide

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– Fig. 1. Basis of the OMA1 activity using fluorescence-based peptide.
Fluorescence is released when OMA1 recognizes and cleaves the OPA1 8-mer
peptide (fluorescence reporter) presumably at the RA site, from the cited paper

The continual fission and fusion the Mitochondria undergoes to change its shape and function are a key trait of the organelle, one that is regulated by the enzyme OMA1. However, there is little known regarding OMA1 due to the lack of a consistent method to measure its activity. More information is needed to truly gauge the role of OMA1 as a therapeutic agent. This is where one group sought to measure this activity utilizing a fluorescence-based reporter cleavage assay, one where the protease OMA1 activity is measured by MCA fluorescent peptide.

OMA1 activity measured by (MCA-AFRATDHG-(lys)DNP) peptide

The group arrived at this specific sequence as it includes the specific point on protein OPA1 (between the arginine and alanine) that OMA1 cleaves. They would then be able to spectrofluorometrically measure the fluorescent MCA moiety after the cleavage takes place. The assay proved successful in measuring the activity of OMA1, and in an inexpensive manner. The work clearly lays out the foundation for future studies of OMA1, in both its normal and abnormal pathology.

Julia Tobacyk, Nirmala Parajuli, Stephen Shrum, John P. Crow, Lee Ann MacMillan-Crow, The first direct activity assay for the mitochondrial protease OMA1, Mitochondrion, Volume 46, 2019, Pages 1-5, ISSN 1567-7249, https://doi.org/10.1016/j.mito.2019.03.001.

Revolutionary Antimicrobial Peptides: A New Hope in the Battle Against Citrus Greening

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Citrus greening, or Huanglongbing (HLB), is a disease that devastates citrus production all over the world. The culprit behind HLB is the bacterium Candidatus Liberibacter spp. (e.g., CLas), an unculturable pathogen that has proven very difficult to treat. Once a tree is infected, it becomes unproductive and dies within years, costing the global citrus market billions. While current attempts to combat HLB rely on controlling the insect vector, scientists have turned some attention toward the potential of peptides. Their work displayed how antimicrobial peptides show promise for combatting citrus greening, mainly by methods against CLas itself.

Antimicrobial peptides effective against CLas bacteria

With not many current effective options to fight HLB, scientists believe the next area of interest is targeting the CLas secretory pathway using antimicrobial peptides provided by LifeTein. Specifically, the antimicrobial peptides would be blocking the TolC efflux pump protein. The study found three peptides capable of doing this by binding tightly with the TolC receptors and even the β barrel entrance of the protein as well. Treatment with peptides in this manner showed effective inhibition and even mortality in models closely resembling CLas.

The studies displayed using antimicrobial peptides show major promise for future treatment of HLB. With the chemical-resistant bacteria CLas being nearly impossible to slow down, peptides just may have been holding the solution all along. There is hope that new therapies can be developed utilizing the strategies shown, and global citrus production can rest easy after decades of HLB ravaging the farms.

Wang, Haoqi, Nirmitee Mulgaonkar, Samavath Mallawarachchi, Manikandan Ramasamy, Carmen S. Padilla, Sonia Irigoyen, Gitta Coaker, Kranthi K. Mandadi, and Sandun Fernando. 2022. “Evaluation of Candidatus Liberibacter Asiaticus Efflux Pump Inhibition by Antimicrobial Peptides” Molecules 27, no. 24: 8729. https://doi.org/10.3390/molecules27248729

Peptides Fold and Self-Assemble on Graphite-Water Interfaces

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J. Chem. Inf. Model. 2022, 62, 17, 4066-4082

The concept of self-assembling peptides is a promising front where construction of devices can be achieved through a single molecule. While the outcome is enticing, the means to reach a consistent outcome are complex to say the least. Dozens of factors go into how a peptide may self-assemble and fold, with the most important being the sequence itself. While this can be handled by careful screening and simulations, the interface at which this folding occurs becomes more important to consider at well. Researchers looked to test how specific peptides fold and self-assemble on graphite-water interfaces, where a number of factors give this method the advantage over doing so in free solution.

Graphite helps peptides self fold into conformations

The group studying this phenomenon claimed that the folded conformations of the peptides were stable over a variety of temperatures when observed over graphite. They point out that it is due to the peptide backbone aligning with the zigzag directions of the graphite plane, thus allowing the conformations to occur more favorably from the intermolecular hydrogen bonds of the molecule. Atomic force microscopy revealed these theories to be true beyond initial simulations as well.

The team believes the design principles displayed in these experiments could be of great use in future iterations of self-assembling peptide engineering. The thermodynamically favored self-assembly with the use of a graphite-water interface shows promise as a medium for even more complex molecular devices in the future, a future LifeTein is looking forward to being a part of.

Justin Legleiter, Ravindra Thakkar, Astrid Velásquez-Silva, Ingrid Miranda-Carvajal, Susan Whitaker, John Tomich, and Jeffrey Comer
Journal of Chemical Information and Modeling 2022 62 (17), 4066-4082
DOI: 10.1021/acs.jcim.2c00419