High Affinity Peptide Inhibitors of the Hepatitis C Virus NS3-4A Protease Refractory to Common Resistant Mutants

Kügler, Jonas; Schmelz, Stefan; Gentzsch, Juliane; Haid, Sibylle; Pollmann, Erik; Heuvel, Joop van den; Franke, Raimo; Pietschmann, Thomas; Heinz, Dirk W.; Collins, John

doi:10.1074/jbc.m112.393843

Cited by 28 publications

(21 citation statements)

References 46 publications

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“…4a). The peptide conformation agrees well with those of related proteasepeptide complexes (Wilmouth et al, 2001;Kü gler et al, 2012;Lechtenberg et al, 2013; PDB entries 1hax, 4bxw and 4a1t) and includes several possible hydrogen bonds between the main chain of LysC and the substrate (Fig. 4a).…”

Section: Lysc Substrate-binding Site and Differences Between Wild-typsupporting

confidence: 71%

“…7) but also in the peptide-bound form in other proteases (model in Fig. 4a; Wilmouth et al, 2001;Kü gler et al, 2012;Lechtenberg et al, 2013), enabling facile transfer of protons to and from the intermediate. The oxyanion hole therefore may not need to stabilize a highly energetic alkoxide with a pK a of >16, but serves to hydrogen-bond both electron lone pairs of the hydroxyl O atom while the third valence is pointing into, and is protonated by, solvent.…”

Section: The Oxyanion Hole In Lysc Can Also Be An 'Alco-hole'mentioning

confidence: 99%

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Atomic resolution structure of a lysine-specific endoproteinase fromLysobacter enzymogenessuggests a hydroxyl group bound to the oxyanion hole

Asztalos

Müller

Hölke

et al. 2014

Acta Cryst D Biol Crystallogr

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Lysobacter enzymogenes lysyl endoproteinase (LysC) is a trypsin-type serine protease with a high pH optimum that hydrolyses all Lys-Xaa peptide bonds. The high specificity of LysC renders it useful for biotechnological purposes. The K30R variant of a related lysyl endoproteinase from Achromobacter lyticus has favourable enzymatic properties that might be transferrable to LysC. To visualize structural differences in the substrate-binding sites, the crystal structures of wild-type and the K30R variant of LysC were determined. The mutation is located at a distance of 12 Å from the catalytic triad and subtly changes the surface properties of the substrate-binding site. The high pH optimum of LysC can be attributed to electrostatic effects of an aromatic Tyr/His stack on the catalytic aspartate and is a general feature of this enzyme subfamily. LysC crystals in complex with the covalent inhibitor N(α)-p-tosyl-lysyl chloromethylketone yielded data to 1.1 and 0.9 Å resolution, resulting in unprecedented precision of the active and substrate-binding sites for this enzyme subfamily. Error estimates on bond lengths and difference electron density indicate that instead of the expected oxyanion a hydroxyl group binds to the partially solvent-exposed oxyanion hole. Protonation of the alkoxide catalytic intermediate might be a recurring feature during serine protease catalysis.

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Section: Lysc Substrate-binding Site and Differences Between Wild-typsupporting

confidence: 71%

Section: The Oxyanion Hole In Lysc Can Also Be An 'Alco-hole'mentioning

confidence: 99%

Atomic resolution structure of a lysine-specific endoproteinase fromLysobacter enzymogenessuggests a hydroxyl group bound to the oxyanion hole

Asztalos

Müller

Hölke

et al. 2014

Acta Cryst D Biol Crystallogr

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“…To provide a third reader, we used a previously described, genetically-encoded peptide that binds to the active site of apo NS3a, here called apo NS3a reader (ANR). 26,27 ANR forms a basal complex with NS3a with an affinity of 10 nM that is disrupted by NS3atargeting drugs (Fig. 1a).…”

Section: Computational Design Of Ns3a Readersmentioning

confidence: 99%

Multi-input chemical control of protein dimerization for programming graded cellular responses

et al. 2019

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Chemical and optogenetic methods for post-translationally controlling protein function have enabled new discoveries and the engineering of synthetic cellular functions. However, most of these methods only confer single-input, single-output control. To increase the diversity of posttranslational behaviors that can be programmed we built a system based on a single protein receiver that can integrate multiple drug inputs, including approved therapeutics. Our system translates drug inputs into diverse outputs with engineered reader proteins that provide variable dimerization states of the receiver protein. We show that our single receiver protein architecture can be used to program diverse cellular responses, including graded and proportional dual-output control of transcription and mammalian cell signaling. We apply our tools to titrate the competing activities of the Rac and Rho GTPases to control cell morphology. Our receiver protein and suite Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:

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“…Protease-inhibiting peptides have been developed against HCV infection. These molecules were conjugated to a selection of CPPs and the fusion with Antennapedia was regarded as the approach with the highest effi ciency [ 138 ]. Peptide-targeting HIV-1 capsid formation has been successfully applied using a commercial CPP-based transfection reagent, Chariot.…”

Section: Cpps As Antiviral Drug Delivery Vectorsmentioning

confidence: 99%

The Antimicrobial and Antiviral Applications of Cell-Penetrating Peptides

Pärn

Eriste

Langel

2015

Methods in Molecular Biology

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Over the past two decades, cell-penetrating peptides (CPPs) have become increasingly popular both in research and in application. There have been numerous studies on the physiochemical characteristics and behavior of CPPs in various environments; likewise, the mechanisms of entry and delivery capabilities of these peptides have also been extensively researched. Besides the fundamental issues, there is an enormous interest in the delivery capabilities of the peptides as the family of CPPs is a promising and mostly non-toxic delivery vector candidate for numerous medical applications such as gene silencing, transgene delivery, and splice correction. Lately, however, there has been an emerging field of study besides the high-profile gene therapy applications-the use of peptides and CPPs to combat various infections caused by harmful bacteria, fungi, and viruses.In this chapter, we aim to provide a short overview of the history and properties of CPPs which is followed by more thorough descriptions of antimicrobial and antiviral peptides. To achieve this, we analyze the origin of such peptides, give an overview of the mechanisms of action and discuss the various practical applications which are ongoing or have been suggested based on research.

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High Affinity Peptide Inhibitors of the Hepatitis C Virus NS3-4A Protease Refractory to Common Resistant Mutants

Cited by 28 publications

References 46 publications

Atomic resolution structure of a lysine-specific endoproteinase fromLysobacter enzymogenessuggests a hydroxyl group bound to the oxyanion hole

Atomic resolution structure of a lysine-specific endoproteinase fromLysobacter enzymogenessuggests a hydroxyl group bound to the oxyanion hole

Multi-input chemical control of protein dimerization for programming graded cellular responses

The Antimicrobial and Antiviral Applications of Cell-Penetrating Peptides

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