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Articles 1 - 9 of 9

Full-Text Articles in Biochemistry, Biophysics, and Structural Biology

The Sh3 Domain Of Unc-89 (Obscurin) Interacts With Paramyosin, A Coiled-Coil Protein, In Caenorhabditis Elegans Muscle, Hiroshi Qadota, Jonathan Mcmurry, Verra M. Ngwa, Et Al. Mar 2017

The Sh3 Domain Of Unc-89 (Obscurin) Interacts With Paramyosin, A Coiled-Coil Protein, In Caenorhabditis Elegans Muscle, Hiroshi Qadota, Jonathan Mcmurry, Verra M. Ngwa, Et Al.

Jonathan McMurry

UNC-89 is a giant polypeptide located at the sarcomeric M-line of Caenorhabditis elegans muscle. The human homologue is obscurin. To understand how UNC-89 is localized and functions, we have been identifying its binding partners. Screening a yeast two-hybrid library revealed that UNC-89 interacts with paramyosin. Paramyosin is an invertebrate-specific coiled-coil dimer protein that is homologous to the rod portion of myosin heavy chains and resides in thick filament cores. Minimally, this interaction requires UNC-89’s SH3 domain and residues 294–376 of paramyosin and has a KD of ∼1.1 μM. In unc-89 loss-of-function mutants that lack the SH3 domain, paramyosin is found …


Localisation And Protein-Protein Interactions Of The Helicobacter Pylori Taxis Sensor T1pd And Their Connection To Metabolic Functions, Wiebke Behrens, Tobias Schweinitzer, Jonathan L. Mcmurry, Christine Josenhans Mar 2017

Localisation And Protein-Protein Interactions Of The Helicobacter Pylori Taxis Sensor T1pd And Their Connection To Metabolic Functions, Wiebke Behrens, Tobias Schweinitzer, Jonathan L. Mcmurry, Christine Josenhans

Jonathan McMurry

The Helicobacter pylori energy sensor TlpD determines tactic behaviour under low energy conditions and is important in vivo. We explored protein-protein interactions of TlpD and their impact on TlpD localisation and function. Pull-down of tagged TlpD identified protein interaction partners of TlpD, which included the chemotaxis histidine kinase CheAY2, the central metabolic enzyme aconitase (AcnB) and the detoxifying enzyme catalase (KatA). We confirmed that KatA and AcnB physically interact with TlpD. While the TlpD-dependent behavioural response appeared not influenced in the interactor mutants katA and acnB in steady-state behavioural assays, acetone carboxylase subunit (acxC) mutant behaviour was altered. TlpD was …


Novel Cell Penetrating Peptide-Adaptors Effect Intracellular Delivery And Endosomal Escape Of Protein Cargos, John C. Salerno, Verra M. Ngwa, Scott J. Nowak, Carol A. Chrestensen, Allison N. Healey, Jonathan L. Mcmurry Mar 2017

Novel Cell Penetrating Peptide-Adaptors Effect Intracellular Delivery And Endosomal Escape Of Protein Cargos, John C. Salerno, Verra M. Ngwa, Scott J. Nowak, Carol A. Chrestensen, Allison N. Healey, Jonathan L. Mcmurry

Jonathan McMurry

The use of cell penetrating peptides (CPPs) as biomolecular delivery vehicles holds great promise for therapeutic and other applications, but development has been stymied by poor delivery and lack of endosomal escape. We have developed a CPP-adaptor system capable of efficient intracellular delivery and endosomal escape of user-defined protein cargos. The cell penetrating sequence of HIV transactivator of transcription was fused to calmodulin, which binds with subnanomolar affinity to proteins containing a calmodulin binding site. Our strategy has tremendous advantage over prior CPP technologies because it utilizes high affinity noncovalent, but reversible coupling between CPP and cargo. Three different cargo …


Flagellar Formation In C-Ring-Defective Mutants By Overproduction Of Flii, The Atpase Specific For Flagellar Type Iii Secretion, Manabu Konishi, Masaomi Kanbe, Jonathan L. Mcmurry, Shin-Ichi Aizawa Mar 2017

Flagellar Formation In C-Ring-Defective Mutants By Overproduction Of Flii, The Atpase Specific For Flagellar Type Iii Secretion, Manabu Konishi, Masaomi Kanbe, Jonathan L. Mcmurry, Shin-Ichi Aizawa

Jonathan McMurry

The flagellar cytoplasmic ring (C ring), which consists of three proteins, FliG, FliM, and FliN, is located on the cytoplasmic side of the flagellum. The C ring is a multifunctional structure necessary for flagellar protein secretion, torque generation, and switching of the rotational direction of the motor. The deletion of any one of the fliG, fliM, and fliN genes results in a Fla - phenotype. Here, we show that the overproduction of the flagellum-specific ATPase FliI overcomes the inability of basal bodies with partial C-ring structures to produce complete flagella. Flagella made upon FliI overproduction were paralyzed, indicating that an …


Identification Of Preferred Dna-Binding Sites For The Thermus Thermophilus Transcriptional Regulator Sbtr By The Combinatorial Approach Repsa, Michael W. Van Dyke, Matthew D. Beyer, Emily Clay, Jonathan L. Mcmurry Mar 2017

Identification Of Preferred Dna-Binding Sites For The Thermus Thermophilus Transcriptional Regulator Sbtr By The Combinatorial Approach Repsa, Michael W. Van Dyke, Matthew D. Beyer, Emily Clay, Jonathan L. Mcmurry

Jonathan McMurry

One of the first steps towards elucidating the biological function of a putative transcriptional regulator is to ascertain its preferred DNA-binding sequences. This may be rapidly and effectively achieved through the application of a combinatorial approach, one involving very large numbers of randomized oligonucleotides and reiterative selection and amplification steps to enrich for high-affinity nucleic acid-binding sequences. Previously, we had developed the novel combinatorial approach Restriction Endonuclease Protection, Selection and Amplification (REPSA), which relies not on the physical separation of ligand-nucleic acid complexes but instead selects on the basis of ligand-dependent inhibition of enzymatic template inactivation, specifically cleavage by type …


Efn-4 Functions In Lad-2-Mediated Axon Guidance In Caenorhabditis Elegans, Alicia A. Schwieterman, Cory J. Donelson, Jonathan L. Mcmurry, Martin L. Hudson Mar 2017

Efn-4 Functions In Lad-2-Mediated Axon Guidance In Caenorhabditis Elegans, Alicia A. Schwieterman, Cory J. Donelson, Jonathan L. Mcmurry, Martin L. Hudson

Jonathan McMurry

During development of the nervous system, growing axons rely on guidance molecules to direct axon pathfinding. A well-characterized family of guidance molecules are the membrane-associated ephrins, which together with their cognate Eph receptors, direct axon navigation in a contact-mediated fashion. InC. elegans, the ephrin-Eph signaling system is conserved and is best characterized for their roles in neuroblast migration during early embryogenesis. This study demonstrates a role for theC. elegansephrin EFN-4 in axon guidance. We provide both genetic and biochemical evidence that is consistent with theC. elegansdivergent L1 cell adhesion molecule LAD-2 acting as a non-canonical ephrin receptor to EFN-4 to …


Characterization Of Myxococcus Xanthus Mazf And Implications For A New Point Of Regulation, Tye O. Boynton, Jonathan L. Mcmurry, Lawrence J. Shimkets Mar 2013

Characterization Of Myxococcus Xanthus Mazf And Implications For A New Point Of Regulation, Tye O. Boynton, Jonathan L. Mcmurry, Lawrence J. Shimkets

Jonathan McMurry

During development, Myxococcus xanthus cells undergo programmed cell death (PCD) whereby 80% of vegetative cells die. Previously, the MazF RNA interferase has been implicated in this role. Recently, it was shown that deletion of the mazF gene does not eliminate PCD in wild-type strain DK1622 as originally seen in DZF1. To clarify the role of MazF, recombinant enzyme was characterized using a highly sensitive assay in the presence and absence of the proposed antitoxin MrpC. In contrast to previous reports that MrpC inhibits MazF activity, the hydrolysis rate was enhanced in a concentration-dependent manner with MrpC or MrpC2, an N-terminally …


Kinetic Characterization Of Salmonella Flik-Flhb Interactions Demonstrates Complexity Of The Type Iii Secretion Substrate-Specificity Switch, Daniel P. Morris, Eric D. Roush, J. Will Thompson, M. Arthur Moseley, James W. Murphy, Jonathan L. Mcmurry Apr 2011

Kinetic Characterization Of Salmonella Flik-Flhb Interactions Demonstrates Complexity Of The Type Iii Secretion Substrate-Specificity Switch, Daniel P. Morris, Eric D. Roush, J. Will Thompson, M. Arthur Moseley, James W. Murphy, Jonathan L. Mcmurry

Jonathan McMurry

The bacterial flagellum is a complex macromolecular machine consisting of more than 20000 proteins, most of which must be exported from the cell via a dedicated Type III secretion apparatus. At a defined point in flagellar morphogenesis, hook completion is sensed and the apparatus switches substrate specificity type from rod and hook proteins to filament ones. How the switch works is a subject of intense interest. FIiK and F1hBs play central roles. In the present study, two optical biosensing methods were used to characterize FIiK-F1hB interactions using wild-type and two variant FlhBs from mutants with severe flagellar structural defects. Binding …


The Helicobacter Pylori Anti-Sigma Factor Flgm Is Predominantly Cytoplasmic And Cooperates With The Flagellar Basal Body Protein Flha, Melanie Rust, Sophie Borchert, Eike Niehus, Sarah A. Gripp, Afrodita Bajceta, Jonathan L. Mcmurry, Sebastian Suerbaum, Kelly T. Hughes, Christine Josenhans Apr 2011

The Helicobacter Pylori Anti-Sigma Factor Flgm Is Predominantly Cytoplasmic And Cooperates With The Flagellar Basal Body Protein Flha, Melanie Rust, Sophie Borchert, Eike Niehus, Sarah A. Gripp, Afrodita Bajceta, Jonathan L. Mcmurry, Sebastian Suerbaum, Kelly T. Hughes, Christine Josenhans

Jonathan McMurry

Helicobacter pylori requires flagellar motility and orientation to persist actively in its habitat. A particular feature of flagella in most Helicobacter species including H. pylori is a membraneous flagellar sheath. The anti-sigma factor FlgM of H. pylori is unusual, since it lacks an N-terminal domain present in other FlgM homologs, e.g., FlgM of Salmonella spp., whose regulatory function is intimately coupled to its secretion through the flagellar type III secretion system. The aim of the present study was to characterize the localization and secretion of the short H. pylori FlgM in the presence of a flagellar sheath and to elucidate …