• Title/Summary/Keyword: effector domain

Search Result 42, Processing Time 0.026 seconds

Evidence of complex formation between FADD and c-FLIP death effector domains for the death inducing signaling complex

  • Hwang, Eun Young;Jeong, Mi Suk;Park, So Young;Jang, Se Bok
    • BMB Reports
    • /
    • v.47 no.9
    • /
    • pp.488-493
    • /
    • 2014
  • Adaptor protein FADD forms the death inducing signaling complex (DISC) by recruiting the initiating caspases-8 and -10 through homotypic death effector domain (DED) interactions. Cellular FLICE-inhibitory protein (c-FLIP) is an inhibitor of death ligand-induced apoptosis downstream of death receptors, and FADD competes with procaspase-8/10 for recruitment for DISC. However, the mechanism of action of FADD and c-FLIP proteins remain poorly understood at the molecular level. In this study, we provide evidence indicating that the death effector domain (DED) of FADD interacts directly with the death effector domain of human c-FLIP. In addition, we use homology modeling to develop a molecular docking model of FADD and c-FLIP proteins. We also find that four structure-based mutants (E80A, L84A, K169A and Y171A) of c-FLIP DEDs disturb the interaction with FADD DED, and that these mutations lower the stability of the c-FLIP DED.

Mutational Analysis of the Effector Domain of Brassica Sar1 Protein

  • Kim, Min-Gab;Lee, Jung-Ro;Lim, Hye-Song;Shin, Mi-Rim;Cheon, Min-Gyeong;Lee, Deok-Ho;Kim, Woe-Yeon;Lee, Sang-Yeol
    • Journal of Applied Biological Chemistry
    • /
    • v.50 no.3
    • /
    • pp.109-114
    • /
    • 2007
  • Sar1p is a ras-related GTP-binding protein that functions in intracellular protein transport between the endoplasmic reticulum (ER) and the Golgi complex. The effector domain of Ras family proteins is highly conserved and this domain is functionally interchangeable in plant, yeast and mammalian Sar1. Using a recombinant Brassica sar1 protein (Bsar1p) harboring point mutations in its effector domain, we here investigated the ability of Sar1p to bind and hydrolyze GTP and to interact with the two sar1-specific regulators, GTPase activating protein (GAP) and guanine exchange factor (GEF). The T51A and T55A mutations impaired Bsar1p intrinsic GTP-binding and GDP-dissociation activity. In contrast, mutations in the switch domain of Bsar1 did not affect its intrinsic GTPase activity. Moreover, the P50A, P54A, and S56A mutations affected the interaction between Bsar1p and GAP. P54A mutant protein did not interact with two regulating proteins, GEF and GAP, even though the mutation didn't affect the intrinsic GTP-binding, nucleotide exchange or GTPase activity of Bsar1p.

Structural insights showing how arginine is able to be glycosylated by pathogenic effector proteins

  • Park, Jun Bae;Yoo, Youngki;Cho, Hyun-Soo
    • BMB Reports
    • /
    • v.51 no.12
    • /
    • pp.609-610
    • /
    • 2018
  • Glycosylation is one form of protein modification and plays a key role in protein stability, function, signaling regulation and even cancer. NleB and SseK are bacterial effector proteins and possess glycosyltransferase activity, even though they have different substrate preferences. NleB/SseKs transfer the GlcNAc sugar to an arginine residue of host proteins, leading to reduced $NF-{\kappa}B-dependent$ responses. By combining X-ray crystallography, NMR, molecular dynamics, enzyme kinetic assays and in vivo experiments, we demonstrated that a conserved HEN (His-Glu-Asn) motif in the active site plays a key role in enzyme catalysis and virulence. The lid-domain regulates the opening and closing of the active site and the HLH domain determines the substrate specificity. Our findings provide evidence for the enzymatic mechanism by which arginine can be glycosylated by SseK/NleB enzymes.

The Role of SH2 Domain-containing Leukocyte Phosphoprotein of 76 kDa in the Regulation of Immune Cell Development and Function

  • Koretzky, Gary A.
    • IMMUNE NETWORK
    • /
    • v.9 no.3
    • /
    • pp.75-83
    • /
    • 2009
  • Recent years have seen an explosion of new knowledge defining the molecular events that are critical for development and activation of immune cells. Much of this new information has come from a careful molecular dissection of key signal transduction pathways that are initiated when immune cell receptors are engaged. In addition to the receptors themselves and critical effector molecules, these signaling pathways depend on adapters, proteins that have no intrinsic effector function but serve instead as scaffolds to nucleate multimolecular complexes. This review summarizes some of what has been learned about one such adapter protein, SH2 domain-containing leukocyte phosphoprotein of 76 kDa (SLP-76), and how it regulates and integrates signals after engagement of immunoreceptors and integrins on various immune cell lineages.

The Effector Functions of Mature T Lymphocytes Are Impaired in Transgenic Mice Expressing the SH2 Domain of TSAd/Lad

  • Choi, Youngbong;Park, Eunkyung;Ahn, Eunseon;Park, Inyoung;Yun, Yungdae
    • Molecules and Cells
    • /
    • v.28 no.3
    • /
    • pp.183-188
    • /
    • 2009
  • TSAd/Lad is a T cell adaptor molecule involved in $p56^{lck}$-mediated T cell activation. To investigate the functions of TSAd in T cells, we generated transgenic (TG) mice expressing the SH2 domain of TSAd (TSAd-SH2) under the control of the $p56^{lck}$ proximal promoter. In T cells from TSAd-SH2 TG mice, T cell receptor (TCR)-mediated early signaling events, such as $Ca^{2+}$ flux and ERK activation, were normal; however, late activation events, such as IL-2 production and proliferation, were significantly reduced. Moreover, TCR-induced cell adhesion to extracellular matrix (ECM) proteins and migration through ECM proteins were defective in T cells from TSAd-SH2 TG mice. Furthermore, the contact hypersensitivity (CHS) reaction, an inflammatory response mainly mediated by T helper 1 (Th1) cells, was inhibited in TSAd-SH2 TG mice. Taken together, these results show that TSAd, particularly the SH2 domain of TSAd, is essential for the effector functions of T cells.

Modulation of Rit Activation by the Alpha Subunit of Go

  • Yang, Chul-Min;Ghil, Sung-Ho
    • Biomedical Science Letters
    • /
    • v.15 no.4
    • /
    • pp.327-333
    • /
    • 2009
  • Heterotrimeric GTP binding proteins, G-proteins, mediate signal transduction generated by neurotransmitters and hormones. Among G-proteins, Go proteins are the most abundant in brain and classified as a member of Gi family. Ras-like protein in all tissues (Rit), one of the small GTPases, is a member of a Ras superfamily and identified as an important regulator of neuronal differentiation and cell transformation. Recently, we have reported that Rit functioned as a candidate downstream effector for alpha subunit of Go proteins ($Go{\alpha}$) and regulated neurite outgrowth triggered by $Go{\alpha}$ activation. In this study, we showed that the GTPase domain of $Go{\alpha}$ contributed to the direct interaction with Rit. We also demonstrated that $Go{\alpha}$ could lead to an increase of Rit activity suggesting that Rit play a role as a downstream effector of $Go{\alpha}$.

  • PDF

Effect of an End-effector Type of Robotic Gait Training on Stand Capability, Locomotor Function, and Gait Speed in Individuals with Spastic Cerebral Palsy (엔드 이펙터 타입의 로봇보행훈련이 뇌성마비인의 서기, 보행 기능과 보행속도에 미치는 영향)

  • Hwang, Jongseok
    • Journal of the Korean Society of Physical Medicine
    • /
    • v.16 no.3
    • /
    • pp.123-130
    • /
    • 2021
  • PURPOSE: Robotic gait training is being used increasingly to improve the gross motor performance and gait speed. The present study examined the effectiveness of a novel end-effector type of robotic gait training (RGT) system on standing, walking, running, and jumping functions, as well as the gait speed in children with spastic cerebral palsy. METHODS: Eleven children with spastic cerebral palsy Gross Motor Function Classification System (GMFCS) levels I-III (6 males; age range, 15.09 ± 1.44 years) were examined. They underwent 24 sessions (30 minutes/sessions, one time/day, three days/week for eight consecutive weeks) of RGT. The Gross Motor Function Measure-88 D domain (GMFM D), and GMFM E were assessed with a pretest and posttest of RGT. The setting was a one-group pretest-posttest design. RESULTS: A comparison of the pre-test and post-test show that the outcomes in post-test of GMFM D (p < .01), GMFM E (p < .05), and 10MWT were improved significantly after RGT intervention. CONCLUSION: The present study provided the first evidence on the effects of an eight-weeks RGT intervention in participants with spastic CP. The outcomes of this clinical study showed that standing performance, locomotion function, and gait speed increased in after 24 sessions of the end-effector RGT system in children with spastic cerebral palsy.

Crystal Structure of the Pneumococcal Vancomycin-Resistance Response Regulator DNA-Binding Domain

  • Park, Sang-Sang;Lee, Sangho;Rhee, Dong-Kwon
    • Molecules and Cells
    • /
    • v.44 no.3
    • /
    • pp.179-185
    • /
    • 2021
  • Vancomycin response regulator (VncR) is a pneumococcal response regulator of the VncRS two-component signal transduction system (TCS) of Streptococcus pneumoniae. VncRS regulates bacterial autolysis and vancomycin resistance. VncR contains two different functional domains, the N-terminal receiver domain and C-terminal effector domain. Here, we investigated VncR C-terminal DNA binding domain (VncRc) structure using a crystallization approach. Crystallization was performed using the micro-batch method. The crystals diffracted to a 1.964 Å resolution and belonged to space group P212121. The crystal unit-cell parameters were a = 25.71 Å, b = 52.97 Å, and c = 60.61 Å. The structure of VncRc had a helix-turn-helix motif highly similar to the response regulator PhoB of Escherichia coli. In isothermal titration calorimetry and size exclusion chromatography results, VncR formed a complex with VncS, a sensor histidine kinase of pneumococcal TCS. Determination of VncR structure will provide insight into the mechanism by how VncR binds to target genes.

Advanced T and Natural Killer Cell Therapy for Glioblastoma

  • Wan-Soo Yoon;Dong-Sup Chung
    • Journal of Korean Neurosurgical Society
    • /
    • v.66 no.4
    • /
    • pp.356-381
    • /
    • 2023
  • Although immunotherapy has been broadly successful in the treatment of hematologic malignancies and a subset of solid tumors, its clinical outcomes for glioblastoma are still inadequate. The results could be due to neuroanatomical structures such as the blood-brain-barrier, antigenic heterogeneity, and the highly immunosuppressive microenvironment of glioblastomas. The antitumor efficacy of endogenously activated effector cells induced by peptide or dendritic cell vaccines in particular has been insufficient to control tumors. Effector cells, such as T cells and natural killer (NK) cells can be expanded rapidly ex vivo and transferred to patients. The identification of neoantigens derived from tumor-specific mutations is expanding the list of tumor-specific antigens for glioblastoma. Moreover, recent advances in gene-editing technologies enable the effector cells to not only have multiple biological functionalities, such as cytokine production, multiple antigen recognition, and increased cell trafficking, but also relieve the immunosuppressive nature of the glioblastoma microenvironment by blocking immune inhibitory molecules, which together improve their cytotoxicity, persistence, and safety. Allogeneic chimeric antigen receptor (CAR) T cells edited to reduce graft-versus-host disease and allorejection, or induced pluripotent stem cell-derived NK cells expressing CARs that use NK-specific signaling domain can be a good candidate for off-the-shelf products of glioblastoma immunotherapy. We here discuss current progress and future directions for T cell and NK cell therapy in glioblastoma.

Engineering and Application of Zinc Finger Proteins and TALEs for Biomedical Research

  • Kim, Moon-Soo;Kini, Anu Ganesh
    • Molecules and Cells
    • /
    • v.40 no.8
    • /
    • pp.533-541
    • /
    • 2017
  • Engineered DNA-binding domains provide a powerful technology for numerous biomedical studies due to their ability to recognize specific DNA sequences. Zinc fingers (ZF) are one of the most common DNA-binding domains and have been extensively studied for a variety of applications, such as gene regulation, genome engineering and diagnostics. Another novel DNA-binding domain known as a transcriptional activator-like effector (TALE) has been more recently discovered, which has a previously undescribed DNA-binding mode. Due to their modular architecture and flexibility, TALEs have been rapidly developed into artificial gene targeting reagents. Here, we describe the methods used to design these DNA-binding proteins and their key applications in biomedical research.