For many applications it would be desirable to able control the activity of proteins by using an external signal. In present study, we have explored possibility modulating a restriction enzyme with light. By cross-linking two suitably located cysteine residues bifunctional azobenzene derivative, which can adopt cis - or trans -configuration when illuminated UV blue light, respectively, enzymatic controlled in reversible manner. To determine cross-linked show largest “photoswitch effect,”...

The gene encoding alkaline phosphatase (AP) from the psychrophilic strain TAB5 was cloned, and its nucleotide sequence determined. A single open reading frame consisting of 1125 base pairs which encodes a polypeptide signal peptide 22 amino acids mature protein 353 identified. deduced AP exhibits 38% identity to III IV sequences Bacillus subtilis conserves typical motifs core structure active sites APs various sources. Based on crystal mutated Escerichia coli D153H, homology‐based 3D model...

Abstract Background The central role of Type III secretion systems (T3SS) in bacteria-plant interactions is well established, yet unexpected findings are being uncovered through bacterial genome sequencing. Some Pseudomonas syringae strains possess an uncharacterized cluster genes encoding putative components a second T3SS (T3SS-2) addition to the characterized Hrc1 which associated with disease lesions host plants and triggering hypersensitive response non-host plants. aim this study...

Type VI secretion systems (T6SS) of Gram-negative bacteria form injectisomes that have the potential to translocate effector proteins into eukaryotic host cells. In silico analysis genomes in six Pseudomonas syringae pathovars revealed P. pv. tomato DC3000, tabaci ATCC 11528, T1 and oryzae 1-6 each carry two putative T6SS gene clusters (HSI-I HSI-II; HSI: Hcp island), whereas phaseolicola 1448A B728 one. The DC3000 HSI-I HSI-II possess a highly similar organization nucleotide sequence, 11528...

Type III secretion systems enable plant and animal bacterial pathogens to deliver virulence proteins into the cytosol of eukaryotic host cells, causing a broad spectrum diseases including bacteremia, septicemia, typhoid fever, bubonic plague in mammals, localized lesions, systemic wilting, blights plants. In addition, type are also required for biogenesis flagellum. The HrcQ B protein, component apparatus Pseudomonas syringae with homologues all systems, has variable N-terminal conserved...

Gene clusters encoding various type III secretion system (T3SS) injectisomes, frequently code downstream of the conserved atpase gene for small hydrophilic proteins whose amino acid sequences display a propensity intrinsic disorder and coiled-coil formation. These properties were confirmed experimentally member this class, HrpO protein from T3SS Pseudomonas syringae pv phaseolicola: exhibits high alpha-helical content with characteristics, strikingly low melting temperature, structural that...

In an effort to explore the role of glycine clusters on cold adaptation enzymes, we designed point mutations aiming alter distribution residues close active site psychrophilic alkaline phosphatase from Antarctic strain TAB5. The mutagenesis targets were Gly261 and Gly262. replacement Gly262 by Ala resulted in inactive enzyme. Substitution enzyme with lower stability increased energy activation. double mutant G261A/Y269A basis side‐chain packing criteria a modelled structure restoration...

It has been proposed that certain type II restriction enzymes (REs), such as EcoRV, track the helical pitch of DNA they diffuse along DNA, a so-called rotation-coupled sliding. As yet, there is no direct experimental observation this phenomenon, but mounting indirect evidence gained from single-molecule imaging RE–DNA complexes support hypothesis. We address issue by conjugating fluorescent labels varying size (organic dyes, proteins and quantum dots) to fusing it engineered Rop protein...

Abstract Ralstonia solanacearum depends on numerous virulence factors, also known as effectors, to promote disease in a wide range of economically important host plants. Although some these effectors have been characterized, none yet shown target the host’s secretion machinery. Here, we used an extended library NLR plant immune receptor integrated domains (IDs), identify new effector targets. The screen uncovered that core RipE1, R. species complex, among other targets, associates with...

In the current work we study, via molecular simulations and experiments, folding stability of proteins from tertiary motif 4-α-helical bundles, a recurrent consisting four amphipathic α-helices packed in parallel or antiparallel fashion. The focus is on role loop region structure properties wild-type Rop (wtRop) RM6 proteins, exploring key factors which can affect them, through all-atom dynamics (MD) supporting by experimental findings. A detailed investigation structural conformational...

Abstract Introduction Mammalian glutamate dehydrogenase (hGDH1 in human cells) interconverts to α‐ketoglutarate and ammonia while reducing NAD(P) NAD(P)H. During primate evolution, humans great apes have acquired hGDH2, an isoenzyme that underwent rapid evolutionary adaptation concomitantly with brain expansion, thereby acquiring unique catalytic regulatory properties permitted its function under conditions inhibitory ancestor hGDH1. Although the 3D‐structures of GDHs, including hGDH1, been...

Exo70B1 is a protein subunit of the exocyst complex with crucial role in variety cell mechanisms, including immune responses against pathogens. The calcium-dependent kinase 5 (CPK5) Arabidopsis thaliana (hereafter Arabidopsis), phosphorylates AtExo70B1 upon functional disruption. We previously reported that, Xanthomonas campestris pv. effector XopP compromises AtExo70B1, while bypassing host's hypersensitive response, way that still unclear. Herein we designed an experimental approach, which...

Bacillus cereus is an opportunistic pathogenic bacterium closely related to anthracis , the causative agent of anthrax in mammals. A significant portion B. chromosomal genes are common including which code for putative virulence and surface proteins. thus provides a convenient model organism studying proteins potentially associated with pathogenicity highly infectious . The zinc‐binding protein Bc ZBP, encoded from bc1534 gene has three homologues exhibits deacetylase activity N ‐acetyl...