Solution NMR characterization of chemokine CXCL8/IL-8 monomer and dimer binding to glycosaminoglycans: structural plasticity mediates differential binding interactions

Structural plasticity plays a major role in determining differential binding of CXCL8 monomer and dimer to glycosaminoglycans (GAGs) and that dimer is the high-affinity GAG ligand. We propose that these properties play important roles in orchestrating in vivo chemokine-mediated neutrophil function.

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Synthesis and 31 P NMR characterization of new low toxic highly sensitive pH probes designed for in vivo acidic pH studies

Bioorganic & Medicinal Chemistry ◽

10.1016/s0968-0896(01)00414-x ◽

2002 ◽

Vol 10 (5) ◽

pp. 1451-1458 ◽

Cited By ~ 16

Author(s):

Sophie Martel ◽

Jean-Louis Clément ◽

Agnès Muller ◽

Marcel Culcasi ◽

Sylvia Pietri

Keyword(s):

Acidic Ph ◽

Highly Sensitive ◽

Nmr Characterization ◽

Low Toxic ◽

Ph Probes

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Identification and Characterization of the PutP Proline Permease That Contributes to In Vivo Survival ofStaphylococcus aureus in Animal Models

Infection and Immunity ◽

10.1128/iai.66.2.567-572.1998 ◽

1998 ◽

Vol 66 (2) ◽

pp. 567-572 ◽

Cited By ~ 55

Author(s):

William R. Schwan ◽

Silvija N. Coulter ◽

Eva Y. W. Ng ◽

Michael H. Langhorne ◽

Heather D. Ritchie ◽

...

Keyword(s):

Animal Models ◽

Reading Frame ◽

High Affinity ◽

Wound Model ◽

Infection Models ◽

Uptake Assay ◽

Entire Open Reading Frame ◽

In Vivo Growth

ABSTRACT Staphylococcus aureus is an important pathogen of humans and other animals, causing bacteremia, abscesses, endocarditis, and other infectious syndromes. A signature-tagged mutagenesis (STM) system was adapted for use in studying the genes required for in vivo survival of S. aureus. An STM library was ultimately created in S. aureus RN6390, with Tn917 being used to create the transposon mutations. Pools of S. aureusRN6390 mutants were screened in mouse abscess, bacteremia, and wound infection models for growth attenuation after in vivo passage. One of the mutants that was identified displayed marked attenuation following large-pool screening in all three animal models, which was confirmed in bacteremia and endocarditis models of infection with a smaller pool of mutants. Sequence analysis of the entire open reading frame showed a 99% identity to the high-affinity proline permease (putP) gene characterized in another strain of S. aureus. In wound and murine abscess infection models, the putP mutant was approximately 10-fold more attenuated than was wild-type strain RN6390. Another S. aureus strain transduced with theputP mutation also displayed an attenuated phenotype after passage in the wound model. A [3H]proline uptake assay showed that less proline was specifically transported into theputP mutant than into strain RN6390. The reduced viability of the bacteria possessing the mutation in the S. aureushigh-affinity proline permease suggests that proline scavenging by the bacteria is important for in vivo growth and proliferation and that analogs of proline may serve as potential antistaphylococcal therapeutic agents.

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Evidence that the TRPV1 S1-S4 Membrane Domain Contributes to Thermosensing

10.1101/711499 ◽

2019 ◽

Cited By ~ 1

Author(s):

Minjoo Kim ◽

Nicholas J. Sisco ◽

Jacob K. Hilton ◽

Camila M. Montano ◽

Manuel A. Castro ◽

...

Keyword(s):

Conformational Changes ◽

Solution Nmr ◽

Membrane Domain ◽

Temperature Dependent ◽

Nmr Characterization ◽

Significant Interest ◽

Pore Domain ◽

Coupling Mechanisms

AbstractSensing and responding to temperature is crucial in biology. The TRPV1 ion channel is a well-studied heat-sensing receptor that is also activated by vanilloid compounds including capsaicin. Despite significant interest, the molecular underpinnings of thermosensing have remained elusive. The TRPV1 S1-S4 membrane domain couples chemical ligand binding to the pore domain during channel gating. However, the role of the S1-S4 domain in thermosensing is unclear. Evaluation of the isolated human TRPV1 S1-S4 domain by solution NMR, Far-UV CD, and intrinsic fluorescence shows that this domain undergoes a non-denaturing temperature-dependent transition with a high thermosensitivity. Further NMR characterization of the temperature-dependent conformational changes suggests the contribution of the S1-S4 domain to thermosensing shares features with known coupling mechanisms between this domain with ligand and pH activation. Taken together, this study shows that the TRPV1 S1-S4 domain contributes to TRPV1 temperature-dependent activation.

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Abstract 2786: Pharmacokinetics and in vitro/in vivo characterization of high-affinity bispecific EGFR/CD16A NK cell engagers for the treatment of EGFR-expressing tumors

10.1158/1538-7445.am2018-2786 ◽

2018 ◽

Author(s):

Michael Kluge ◽

Michael Tesar ◽

Uwe Reusch ◽

Stefan Knackmuss ◽

Torsten Haneke ◽

...

Keyword(s):

Nk Cell ◽

High Affinity ◽

In Vivo Characterization

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Approaches to the Solution NMR Characterization of Active Sites for 65 kDa Tetrameric Hemoglobins in the Paramagnetic Cyanomet State

Journal of the American Chemical Society ◽

10.1021/ja9722133 ◽

1997 ◽

Vol 119 (51) ◽

pp. 12643-12654 ◽

Cited By ~ 8

Author(s):

Urszula Kolczak ◽

Chang Han ◽

L. A. Sylvia ◽

Gerd N. La Mar

Keyword(s):

Active Sites ◽

Solution Nmr ◽

Nmr Characterization

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TCR-based therapy for multiple myeloma and other B-cell malignancies targeting intracellular transcription factor BOB1

Blood ◽

10.1182/blood-2016-09-737536 ◽

2017 ◽

Vol 129 (10) ◽

pp. 1284-1295 ◽

Cited By ~ 17

Author(s):

Lorenz Jahn ◽

Pleun Hombrink ◽

Renate S. Hagedoorn ◽

Michel G. D. Kester ◽

Dirk M. van der Steen ◽

...

Keyword(s):

Multiple Myeloma ◽

Transcription Factor ◽

B Cell ◽

B Cell Malignancies ◽

High Affinity ◽

Isolation And Characterization ◽

Key Points

Key Points Isolation and characterization of a high-affinity TCR targeting the intracellular B cell–specific transcription factor BOB1. T cells expressing a BOB1-specific TCR lysed and eradicated primary multiple myeloma and other B-cell malignancies in vitro and in vivo.

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In vivo characterization of the properties of SUMO1-specific monobodies

Biochemical Journal ◽

10.1042/bj20130241 ◽

2013 ◽

Vol 456 (3) ◽

pp. 385-395 ◽

Cited By ~ 4

Author(s):

Anja Berndt ◽

Kevin A. Wilkinson ◽

Michaela J. Heimann ◽

Paul Bishop ◽

Jeremy M. Henley

Keyword(s):

Cost Effective ◽

Effective Alternative ◽

High Affinity ◽

Cost Effective Alternative ◽

In Vivo Characterization ◽

Protein Sumoylation

We demonstrate that SUMO1 monobodies are effective tools for characterizing protein SUMOylation in vivo, and provide a high-affinity, specific and cost-effective alternative to conventional antibodies.

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The GTPase-activating protein ARAP3 regulates chemotaxis and adhesion-dependent processes in neutrophils

Blood ◽

10.1182/blood-2010-10-312959 ◽

2011 ◽

Vol 118 (4) ◽

pp. 1087-1098 ◽

Cited By ~ 40

Author(s):

Laure Gambardella ◽

Karen E. Anderson ◽

Claudia Nussbaum ◽

Anne Segonds-Pichon ◽

Tânia Margarido ◽

...

Keyword(s):

Neutrophil Function ◽

Fine Tuning ◽

Quiescent State ◽

Gtpase Activating Protein ◽

Β2 Integrin ◽

Molecular Components

Abstract Neutrophils form a vital part of the innate immune response, but at the same time their inappropriate activation contributes to autoimmune diseases. Many molecular components are involved in fine-tuning neutrophil function. We report here the first characterization of the role of ARAP3, a PI3K and Rap-regulated GTPase-activating protein for RhoA and Arf6 in murine neutrophils. We show that neutrophils lacking ARAP3 are preactivated in vitro and in vivo, exhibiting increased β2 integrin affinity and avidity. ARAP3-deficient neutrophils are hyperresponsive in several adhesion-dependent situations in vitro, including the formation of reactive oxygen species, adhesion, spreading, and granule release. ARAP3-deficient cells adhere more firmly under flow conditions in vitro and to the vessel wall in vivo. Finally, loss of ARAP3 interferes with integrin-dependent neutrophil chemotaxis. The results of the present study suggest an important function of ARAP3 downstream of Rap. By modulating β2 integrin activity, ARAP3 guards neutrophils in their quiescent state unless activated.

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