• Title/Summary/Keyword: backbone assignment

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Structural Assignment of a Type II PHA Synthase and an Insight Into Its Catalytic Mechanism Using Human Gastric Lipase as the Modeling Template

  • Khairudin, Nurul Bahiyah Ahmad;Samian, Mohd Razip;Najimudin, Nazalan;Wahab, Habibah A
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.173-182
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    • 2005
  • A three dimensional (3D) model for the catalytic region of Type II Pseudomonas sp. USM 4-55 PHA synthase 1 (PhaC1$_{P.sp\;USM\;4-55}$) from residue 267 to residue 484 was developed. Sequence analysis demonstrated that PhaC1$_{P.sp\;USM\;4-55}$ lacked homology with all known structural databases. PSI-BLAST and HMM Superfamily analyses demonstrated that this enzyme belongs to the ${\alpha}/{\beta}$ hydrolase fold family. Threading approach revealed that the most suitable template to use was the Human gastric lipase (1HLG). The superimposition of the predicted PhaC1$_{P.sp\;USM\;4-55}$ model with the 1HLG template structure covering 86.2% of the backbone atoms showed an RMSD of 1.15 ${\AA}$ The catalytic residues comprising of Cys296, Asp451, His452 and His479 were found to be conserved and were located adjacent to each other. We proposed that the catalytic mechanism involved the formation of two tetrahedral intermediates.

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Mainchain NMR Assignments and secondary structure prediction of the C-terminal domain of BldD, a developmental transcriptional regulator from Streptomyces coelicolor A3(2)

  • Kim, Jeong-Mok;Won, Hyung-Sik;Kang, Sa-Ouk
    • Journal of the Korean Magnetic Resonance Society
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    • v.17 no.1
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    • pp.59-66
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    • 2013
  • BldD, a developmental transcription factor from Streptomyces coelicolor, is a homodimeric, DNA-binding protein with 167 amino acids in each subunit. Each monomer consists of two structurally distinct domains, the N-terminal domain (BldD-NTD) responsible for DNA-binding and dimerization and the C-terminal domain (BldD-CTD). In contrast to the BldD-NTD, of which crystal structure has been solved, the BldD-CTD has been characterized neither in structure nor in function. Thus, in terms of structural genomics, structural study of the BldD-CTD has been conducted in solution, and in the present work, mainchain NMR assignments of the recombinant BldD-CTD (residues 80-167 of BldD) could be achieved by a series of heteronuclear multidimensional NMR experiments on a [$^{13}C/^{15}N$]-enriched protein sample. Finally, the secondary structure prediction by CSI and TALOS+ analysis using the assigned chemical shifts data identified a ${\beta}-{\alpha}-{\alpha}-{\beta}-{\alpha}-{\alpha}-{\alpha}$ topology of the domain. The results will provide the most fundamental data for more detailed approach to the atomic structure of the BldD-CTD, which would be essential for entire understanding of the molecular function of BldD.

pH Effect on the Structure of Reduced NifU-like Protein from Helicobacter pylori

  • Lee, Ki-Young;Kim, Ji-Hun;Bae, Ye-Ji;Lee, Bong-Jin
    • Journal of the Korean Magnetic Resonance Society
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    • v.19 no.3
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    • pp.106-111
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    • 2015
  • Helicobacter pylori (H. pylori) survives in acidic and fluctuating pH conditions of the stomach. The pH effect on H. pylori proteins is important for the advanced understanding of its evolution and viability, although this bacterium has the molecular machinery that neutralizes the acidic condition. HP1492 is known as a conserved NifU-like protein from H. pylori. NifU is a nitrogen fixation protein that mediates the transfer of iron-sulfur (Fe-S) cluster to iron-sulfur proteins like ferredoxin. Commonly, the monomeric reduced state of NifU can be converted to the dimeric oxidized state by intermolecular disulfide bond formation. Because it remains unclear that HP1492 actually behaves as known NifU protein, we first found that this protein can adopt both oxidized and reduced forms using size exclusion chromatography. Circular dichroism experiment showed that HP1492 is relatively well-structured at pH 6.5, compared to other pH conditions. On the basis of the backbone resonance assignment of HP1492, we further characterized the residues that are sensitive to pH using NMR spectroscopy. These residues showing large chemical shift changes could be mapped onto the secondary structure of the protein. Our results could provide the foundation for structural and biophysical studies on a wide spectrum of NifU proteins.

Memory Propagation-based Target-aware Segmentation Tracker with Adaptive Mask-attention Decision Network

  • Huanlong Zhang;Weiqiang Fu;Bin Zhou;Keyan Zhou;Xiangbo Yang;Shanfeng Liu
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.18 no.9
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    • pp.2605-2625
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    • 2024
  • Siamese-based segmentation and tracking algorithms improve accuracy and stability for video object segmentation and tracking tasks simultaneously. Although effective, variability in target appearance and background clutter can still affect segmentation accuracy and further influence the performance of tracking. In this paper, we present a memory propagation-based target-aware and mask-attention decision network for robust object segmentation and tracking. Firstly, a mask propagation-based attention module (MPAM) is constructed to explore the inherent correlation among image frames, which can mine mask information of the historical frames. By retrieving a memory bank (MB) that stores features and binary masks of historical frames, target attention maps are generated to highlight the target region on backbone features, thus suppressing the adverse effects of background clutter. Secondly, an attention refinement pathway (ARP) is designed to further refine the segmentation profile in the process of mask generation. A lightweight attention mechanism is introduced to calculate the weight of low-level features, paying more attention to low-level features sensitive to edge detail so as to obtain segmentation results. Finally, a mask fusion mechanism (MFM) is proposed to enhance the accuracy of the mask. By utilizing a mask quality assessment decision network, the corresponding quality scores of the "initial mask" and the "previous mask" can be obtained adaptively, thus achieving the assignment of weights and the fusion of masks. Therefore, the final mask enjoys higher accuracy and stability. Experimental results on multiple benchmarks demonstrate that our algorithm performs outstanding performance in a variety of challenging tracking tasks.