Pawar, Atul

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Pawar
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Atul
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Competitive Metabolite Profiling of Natural Products based on Syringolin A probe.

2021, Pawar, Atul

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Competitive Metabolite Profiling of Natural Products Reveals Subunit Specific Inhibitors of the 20S Proteasome

2020-02-26, Pawar, Atul, Basler, Michael, Goebel, Heike, Alvarez Salinas, Gerardo Omar, Gröttrup, Marcus, Böttcher, Thomas

We have developed a syringolin-based chemical probe and explored its utility for the profiling of metabolite extracts as potent inhibitors of the 20S proteasome. Activity-guided fractionation by competitive labeling allowed us to isolate and identify glidobactin A and C as well as luminmycin A from a Burkholderiales strain. The natural products exhibited unique subunit specificities for the proteolytic subunits of human and mouse constitutive and immunoproteasome in the lower nanomolar range. In particular, glidobactin C displayed an unprecedented β2/β5 coinhibition profile with single-digit nanomolar potency in combination with sufficiently high cell permeability. These properties render glidobactin C a promising live cell proteasome inhibitor with potent activity against human breast cancer cell lines and comparably low immunotoxicity.

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Pseudo-Biomineralization : Complex Mineral Structures Shaped by Microbes

2019-10-14, Durak, Grazyna Malgorzata, Laumann, Michael, Wolf, Stefan L. P., Pawar, Atul, Gebauer, Denis, Böttcher, Thomas

Biomineralization is an active, biologically governed process of mineral formation, established early on in the history of life. The appearance of biomineralizing organisms heavily influenced the course of evolution, leading to the development of the large diversity of the extant taxa. Yet, we are still only beginning to grasp the intricate, genetically regulated mechanisms involved. Since prokaryotic organisms were the first to emerge from the primordial environments, we investigated bacteria–mineral interactions using titration and gas diffusion systems adapted to emulate conditions, which may have facilitated the development of biomineralization initially. By screening the minerals and bacteria from titration experiments with scanning electron microscopy, we discovered a broad spectrum of behavioral strategies employed by bacteria confronted with calcification, which fell into three main categories: (1) evasion of mineralization by the formation of the biofilm, (2) random embedding into the mineral, and (3) control over the mineral shape during its formation. The latter phenomenon we termed pseudo-biomineralization. Our experiments indicate that pseudo-biomineralization is an active process obligatorily reliant on the external calcifying conditions and allowing considerable degree of control over mineral shape, thus producing structures reminiscent of true biominerals. Here, we describe this notion for the first time, thus providing vital insight into the genesis of a transitional stage to calcium carbonate-based biomineralization systems.