Structural and functional characterization of human apolipoprotein E 72-166 peptides in both aqueous and lipid environments

Hsieh, Yi-Hui; Chou, Chi-Yuan

doi:10.1186/1423-0127-18-4

Cited by 18 publications

(10 citation statements)

References 45 publications

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“…Therefore, we developed a two-domain model for the monomeric subunit based on the proposed structures for the WT [2,52], which agrees with our experimental CCS restraints. To build our coarse-grained model, we started with a computational model from Hsieh [52] featuring two distinct domains, in contrast to the NMR structure of apoE3MM where the domains are intermeshed [14] (note also that the mutant, designed to be a monomer, has a considerably different structure that the WTs). The 2-domain structure was split at residue 183 to make a two-body structure, and the CCS of each domain was computed by using the scaled projection approximation function in IMPACT.…”

Section: Coarse-grained Model Of Apoe Tetramersupporting

confidence: 62%

Native Mass Spectrometry, Ion Mobility, Electron-Capture Dissociation, and Modeling Provide Structural Information for Gas-Phase Apolipoprotein E Oligomers

Wang

Eschweiler

Cui

et al. 2019

J. Am. Soc. Mass Spectrom.

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Apolipoprotein E (apoE) is an essential protein in lipid and cholesterol metabolism. Although the three common isoforms in humans differ only at two sites, their consequences in Alzheimer's disease (AD) are dramatically different: only the ε4 allele is a major genetic risk factor for lateonset Alzheimer's disease. The isoforms exist as a mixture of oligomers, primarily tetramer, at low μM concentrations in a lipid-free environment. This self-association is involved in equilibrium with the lipid-free state, and the oligomerization interface overlaps with lipid-binding region. Elucidation of apoE wild-type (WT) structures at an oligomeric state, however, has not yet been achieved. To address this need, we used native electrospray ionization and mass spectrometry (native MS) coupled with ion mobility (IM) to exam the monomer and tetramer of the three WT isoforms. Although collision-induced unfolding (CIU) cannot distinguish the isoforms, the monomeric mutant (MM) of apoE3 shows higher stability when submitted to CIU than the WT monomer. From ion-mobility measurements, we obtained the collision cross section and built a coarse-grained model for the tetramer. Application of electron-capture dissociation (ECD) to the tetramer causes unfolding starting from the C-terminal domain, in good agreement with solution denaturation data and provides additional support for the C4 symmetry structure of the tetramer.

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Section: Coarse-grained Model Of Apoe Tetramersupporting

confidence: 62%

Native Mass Spectrometry, Ion Mobility, Electron-Capture Dissociation, and Modeling Provide Structural Information for Gas-Phase Apolipoprotein E Oligomers

Wang

Eschweiler

Cui

et al. 2019

J. Am. Soc. Mass Spectrom.

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“…The AUC experiments were performed on a XL-A analytical ultracentrifuge (Beckman Coulter) using an An-50 Ti rotor [12,[19][20][21][22]. The sedimentation velocity experiments were performed using a double-sector epon charcoal-filled centerpiece at 20°C with a rotor speed of 42,000 rpm.…”

Section: Analytical Ultracentrifugation Analysismentioning

confidence: 99%

Structural and functional characterization of MERS coronavirus papain-like protease

Lin¹,

Chuang²,

Chen³

et al. 2014

J Biomed Sci

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BackgroundsA new highly pathogenic human coronavirus (CoV), Middle East respiratory syndrome coronavirus (MERS-CoV), has emerged in Jeddah and Saudi Arabia and quickly spread to some European countries since September 2012. Until 15 May 2014, it has infected at least 572 people with a fatality rate of about 30% globally. Studies to understand the virus and to develop antiviral drugs or therapy are necessary and urgent. In the present study, MERS-CoV papain-like protease (PLpro) is expressed, and its structural and functional consequences are elucidated.ResultsCircular dichroism and Tyr/Trp fluorescence analyses indicated that the secondary and tertiary structure of MERS-CoV PLpro is well organized and folded. Analytical ultracentrifugation analyses demonstrated that MERS-CoV PLpro is a monomer in solution. The steady-state kinetic and deubiquitination activity assays indicated that MERS-CoV PLpro exhibits potent deubiquitination activity but lower proteolytic activity, compared with SARS-CoV PLpro. A natural mutation, Leu105, is the major reason for this difference.ConclusionsOverall, MERS-CoV PLpro bound by an endogenous metal ion shows a folded structure and potent proteolytic and deubiquitination activity. These findings provide important insights into the structural and functional properties of coronaviral PLpro family, which is applicable to develop strategies inhibiting PLpro against highly pathogenic coronaviruses.

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“…The AUC experiments were performed on a XL-A analytical ultracentrifuge (Beckman, Fullerton, CA) using an An-50 Ti rotor [15,[17][18][19][20]. The sedimentation velocity experiments were performed using a double-sector epon charcoal-filled centerpiece at 20°C with a rotor speed of 42,000 rpm.…”

Section: Analytical Ultracentrifugation Analysismentioning

confidence: 99%

Differential domain structure stability of the severe acute respiratory syndrome coronavirus papain-like protease

Chou¹,

Cheng²,

Lai³

et al. 2012

Archives of Biochemistry and Biophysics

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Papain-like protease (PLpro) from severe acute respiratory syndrome (SARS) coronavirus is one of the two proteases involved in the proteolytic processing of the virion polyproteins. In addition, PLpro shows significant in vitro deubiquitinating and de-ISGylating activities. All these findings demonstrated the multifunctional nature of the PLpro. Here we report the sensitivity of PLpro to denaturant urea. An increase in urea concentration induced a reversible biphasic unfolding of the enzyme. Differently, the unfolding of the catalytic triad region located within the palm and thumb domains followed a monophasic unfolding curve. Further observations suggest that the zinc-binding domain may start to unfold during the first transition. An 80% lost of its enzymatic activity at a urea concentration lower than 1M showed a close correlation with unfolding of the zinc-binding domain. The enzyme was also characterized in terms of hydrophobicity and size-and-shape distribution. We have demonstrated that PLpro displayed differential domain structure stability and molten globule state in its folding. These studies will not only assist in our understanding of the folding of this viral enzyme, but also that of other deubiquitinating enzymes with a similar scaffold.

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Structural and functional characterization of human apolipoprotein E 72-166 peptides in both aqueous and lipid environments

Cited by 18 publications

References 45 publications

Native Mass Spectrometry, Ion Mobility, Electron-Capture Dissociation, and Modeling Provide Structural Information for Gas-Phase Apolipoprotein E Oligomers

Native Mass Spectrometry, Ion Mobility, Electron-Capture Dissociation, and Modeling Provide Structural Information for Gas-Phase Apolipoprotein E Oligomers

Structural and functional characterization of MERS coronavirus papain-like protease

Differential domain structure stability of the severe acute respiratory syndrome coronavirus papain-like protease

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