Reversible Unfolding of the Severe Acute Respiratory Syndrome Coronavirus Main Protease in Guanidinium Chloride

Chang, Hui-Ping; Chou, Chi-Yuan; Chang, Gu‐Gang

doi:10.1529/biophysj.106.091736

Cited by 39 publications

(68 citation statements)

References 36 publications

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“…5C-H). Before the second unfolding phase (1-3.5 M urea), the broad f/f 0 distribution of the major species suggested a tendency to elongate among the unfolding process, which is similar to that of the SARS-CoV main protease in guanidinium chloride [14]. 3, triangles).…”

Section: Exposure Of the Hydrophobic Region Of Sars-cov Plpro During mentioning

confidence: 60%

“…The thumb, palm, and fingers domains are colored by cyan, green, and orange, respectively. Changes in the maximal peak, intensity, and average emission wavelength (AEW) were used to monitor the unfolding process [14]. The gray sphere is zinc.…”

Section: Spectrofluorimetric Analysismentioning

confidence: 99%

“…The fluorescent dye, 1-anilino-8-naphthalensulfonic acid (ANS), was used as the hydrophobic exposure detecting probe [14][15][16].…”

Section: Ans Spectrofluorimetric Analysismentioning

confidence: 99%

“…Multiple scans at different time intervals were then fitted to a continuous c(s, f/f 0 ) distribution model using the SEDFIT program [14,[21][22][23]. The protein concentrations of 13 lM in 50 mM phosphate buffer at various concentrations of urea.…”

Section: Analytical Ultracentrifugation Analysismentioning

confidence: 99%

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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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Section: Exposure Of the Hydrophobic Region Of Sars-cov Plpro During mentioning

confidence: 60%

Section: Spectrofluorimetric Analysismentioning

confidence: 99%

“…The fluorescent dye, 1-anilino-8-naphthalensulfonic acid (ANS), was used as the hydrophobic exposure detecting probe [14][15][16].…”

Section: Ans Spectrofluorimetric Analysismentioning

confidence: 99%

Section: Analytical Ultracentrifugation Analysismentioning

confidence: 99%

See 2 more Smart Citations

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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“…Each subunit contains a catalytic dyad defined by His41 and Cys145 in the active site (Fig. However, the enzyme is catalytically compe-tent only after dimer formation [9][10][11][12][13][14][15][16][17][18][19][20]. However, the enzyme is catalytically compe-tent only after dimer formation [9][10][11][12][13][14][15][16][17][18][19][20].…”

mentioning

confidence: 99%

Correlation between dissociation and catalysis of SARS-CoV main protease

Lin¹,

Chou²,

Chang³

et al. 2008

Archives of Biochemistry and Biophysics

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The dimeric interface of severe acute respiratory syndrome coronavirus main protease is a potential target for the anti-SARS drug development. We have generated C-terminal truncated mutants by serial truncations. The quaternary structure of the enzyme was analyzed using both sedimentation velocity and sedimentation equilibrium analytical ultracentrifugation. Global analysis of the combined results showed that truncation of C-terminus from 306 to 300 had no appreciable effect on the quaternary structure, and the enzyme remained catalytically active. However, further deletion of Gln-299 or Arg-298 drastically decreased the enzyme activity to 1-2% of wild type (WT), and the major form was a monomeric one. Detailed analysis of the point mutants of these two amino acid residues and their nearby hydrogen bond partner Ser-123 and Ser-139 revealed a strong correlation between the enzyme activity loss and dimer dissociation.

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Molecular Biology of the SARS-Coronavirus

Lal

2010

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The use of general descriptive names, registered names, trademarks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use.Cover Illustration: 3D model of SARS-CoV, with a wedge cut out of it to reveal the nucleocapsid (see Chap. 3 by Daniel R. Beniac and Timothy F. Booth)Background: HL-CZ cells transfected with SARS-CoV Spike construct and incubated with anti-Spike human monoclonal antibody followed by secondary FITC-labeled anti-human antibody (see Chap. 18 by T. Narasaraju, P.L. Soong, J. ter Meulen, J. Goudsmit and Vincent T.K. Chow)Cover design: WMXDesign GmbH, Heidelberg, GermanyPrinted on acid-free paper Springer is part of Springer Science+Business Media (www.springer.com) Foreword SARS was the first new plague of the twenty-first century. Within months, it spread worldwide from its "birthplace" in Guangdong Province, China, affecting over 8,000 people in 25 countries and territories across five continents. SARS exposed the vulnerability of our modern globalised world to the spread of a new emerging infection. SARS (or a similar new emerging disease) could neither have spread so rapidly nor had such a great global impact even 50 years ago, and arguably, it was itself a product of our global inter-connectedness. Increasing affluence and a demand for wild-game as exotic food led to the development of large trade of live animal and game animal markets where many species of wild and domestic animals were co-housed, providing the ideal opportunities for inter-species transmission of viruses and other microbes. Once such a virus jumped species and attacked humans, the increased human mobility allowed the virus the opportunity for rapid spread. An infected patient from Guangdong who stayed for one day at a hotel in Hong Kong led to the transmission of the disease to 16 other guests who travelled on to seed outbreaks of the disease in Toronto, Singapore, and Vietnam, as well as within Hong Kong itself. The virus exploited the practices used in modern intensive care of patients with severe respiratory disease and the weakness in infection control practices within our health care systems to cause outbreaks within hospitals, further amplifying the spread of the disease. Health-care itself has become a two-edged sword.While SARS exposed the vulnerabilities of the modern human condition, it also highlighted the global capacity for a rapid public health and scientific response to an emerging infectious disease threat. Public health and scientific responses succeeded in identifying the causative agent, developing diagnostic tests, and interrupting the spread of the outbreak. The complete virus genome was fully deciphered within weeks and in the ensuing months and years saw an outpouring of scientific research about the disease and its causative agent, the SARS coronavirus. The natural animal reservoir (bats) and amplifier hosts were defined, the virus receptor on human cell...

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Reversible Unfolding of the Severe Acute Respiratory Syndrome Coronavirus Main Protease in Guanidinium Chloride

Cited by 39 publications

References 36 publications

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

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

Correlation between dissociation and catalysis of SARS-CoV main protease

Molecular Biology of the SARS-Coronavirus

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