DIY Methods 2022 Conference Proceedings

As the past years have proven, the methods for conducting and distributing research that we’ve inherited from our disciplinary traditions can be remarkably brittle in the face of rapidly changing social and mobility norms. The ways we work and the ways we meet are questions newly opened for practical and theoretical inquiry; we both need to solve real problems in our daily lives and account for the constitutive effects of these solutions on the character of the knowledge we produce. Methods are not neutral tools, and nor are they fixed ones. As such, the work of inventing, repairing, and hacking methods is a necessary, if often underexplored, part of the wider research process. This conference aims to better interrogate and celebrate such experiments with method. Borrowing from the spirit and circuits of exchange in earlier DIY cultures, it takes the form of a zine ring distributed via postal mail. Participants will craft zines describing methodological experiments and/or how-to guides, which the conference organisers will subsequently mail out to all participants. Feedback on conference proceedings will also proceed through the mail, as well as via an optional Twitter hashtag. The conference itself is thus an experiment with different temporalities and medialities of research exchange. As a practical benefit, this format guarantees that the experience will be free of Zoom fatigue, timezone difficulties, travel expenses, and visa headaches. More generatively, it may also afford slower thinking, richer aesthetic possibilities, more diverse forms of circulation, and perhaps even some amount of delight. The conference format itself is part of the DIY experiment

Profiling constitutive proteolytic events in vivo

Most known organisms encode proteases that are crucial for constitutive proteolytic events. In the present paper, we describe a method to define these events in proteomes from Escherichia coli to humans. The method takes advantage of specific N-terminal biotinylation of protein samples, followed by affinity enrichment and conventional LC (liquid chromatography)–MS/MS (tandem mass spectrometry) analysis. The method is simple, uses conventional and easily obtainable reagents, and is applicable to most proteomics facilities. As proof of principle, we demonstrate profiles of proteolytic events that reveal exquisite in vivo specificity of methionine aminopeptidase in E. coli and unexpected processing of mitochondrial transit peptides in yeast, mouse and human samples. Taken together, our results demonstrate how to rapidly distinguish real proteolysis that occurs in vivo from the predictions based on in vitro experiments

Icatibant , the bradykinin B2 receptor antagonist with target to the interconnected kinin systems

INTRODUCTION: HOE-140/ Icatibant is a selective, competitive antagonist to bradykinin (BK) against its binding to the kinin B2 receptor. Substitution of five non-proteogeneic amino acid analogues makes icatibant resistant to degradation by metalloproteases of kinin catabolism. Icatibant has clinical applications in inflammatory and vascular leakage conditions caused by an acute (non-controlled) production of kinins and their accumulation at the endothelium B2 receptor. The clinical manifestation of vascular leakage, called angioedema (AE), is characterized by edematous attacks of subcutaneous and submucosal tissues, which can cause painful intestinal consequences, and life-threatening complications if affecting the larynx. Icatibant is registered for the treatment of acute attacks of the hereditary BK-mediated AE, i.e., AE due to C1 inhibitor deficiency. AREAS COVERED: This review discusses emerging knowledge on the kinin system: kinin pharmacological properties, biochemical characteristics of the contact phase and kinin catabolism proteases. It underlines the responsibility of the kinins in AE initiation and the potency of icatibant to inhibit AE formation by kinin-receptor interactions. EXPERT OPINION: Icatibant antagonist properties protect BK-mediated AE patients against severe attacks, and could be developed for use in inflammatory conditions. More studies are required to confirm whether or not prolonged and frequent applications of icatibant could result in the impairment of the cardioprotective effect of BK