Biochemical Study of Fibrinolytic Protease from Euphausia superba Possessing Multifunctional Serine Protease Activity
Background: Fibrinolytic protease from Euphausia superba (EFP) was isolated. Objective: Biochemical distinctions, regulation of the catalytic function, and the key residues of EFP were investigated. Methods: The serial inhibition kinetic evaluations coupled with measurements of fluorescence spectra...
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Online Access: | http://dx.doi.org/10.2174/0929866527666201112123714 https://eurekaselect.com/article/download/187909 https://www.eurekaselect.com/187909/article |
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crbenthamsciepub:10.2174/0929866527666201112123714 2024-06-23T07:52:34+00:00 Biochemical Study of Fibrinolytic Protease from Euphausia superba Possessing Multifunctional Serine Protease Activity Qian, Guo-Ying Lim, Gyutae Yin, Shang-Jun Yang, Jun-Mo Lee, Jinhyuk Park, Yong-Doo 2021 http://dx.doi.org/10.2174/0929866527666201112123714 https://eurekaselect.com/article/download/187909 https://www.eurekaselect.com/187909/article en eng Bentham Science Publishers Ltd. Protein & Peptide Letters volume 28, issue 6, page 651-664 ISSN 0929-8665 journal-article 2021 crbenthamsciepub https://doi.org/10.2174/0929866527666201112123714 2024-05-24T13:05:23Z Background: Fibrinolytic protease from Euphausia superba (EFP) was isolated. Objective: Biochemical distinctions, regulation of the catalytic function, and the key residues of EFP were investigated. Methods: The serial inhibition kinetic evaluations coupled with measurements of fluorescence spectra in the presence of 4-(2-aminoethyl) benzene sulfonyl fluoride hydrochloride (AEBSF) was conducted. The computational molecular dynamics (MD) simulations were also applied for a comparative study. Results: The enzyme behaved as a monomeric protein with a molecular mass of about 28.6 kD with K m BApNA = 0.629 ± 0.02 mM and k cat /K m BApNA = 7.08 s-1/mM. The real-time interval measurements revealed that the inactivation was a first-order reaction, with the kinetic processes shifting from a monophase to a biphase. Measurements of fluorescence spectra showed that serine residue modification by AEBSF directly caused conspicuous changes of the tertiary structures and exposed hydrophobic surfaces. Some osmolytes were applied to find protective roles. These results confirmed that the active region of EFP is more flexible than the overall enzyme molecule and serine, as the key residue, is associated with the regional unfolding of EFP in addition to its catalytic role. The MD simulations were supportive to the kinetics data. Conclusion: Our study indicated that EFP has an essential serine residue for its catalyst function and associated folding behaviors. Also, the functional role of osmolytes such as proline and glycine that may play a role in defense mechanisms from environmental adaptation in a krill’s body was suggested. Article in Journal/Newspaper Euphausia superba Bentham Science Publishers Protein & Peptide Letters 28 6 651 664 |
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Bentham Science Publishers |
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crbenthamsciepub |
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English |
description |
Background: Fibrinolytic protease from Euphausia superba (EFP) was isolated. Objective: Biochemical distinctions, regulation of the catalytic function, and the key residues of EFP were investigated. Methods: The serial inhibition kinetic evaluations coupled with measurements of fluorescence spectra in the presence of 4-(2-aminoethyl) benzene sulfonyl fluoride hydrochloride (AEBSF) was conducted. The computational molecular dynamics (MD) simulations were also applied for a comparative study. Results: The enzyme behaved as a monomeric protein with a molecular mass of about 28.6 kD with K m BApNA = 0.629 ± 0.02 mM and k cat /K m BApNA = 7.08 s-1/mM. The real-time interval measurements revealed that the inactivation was a first-order reaction, with the kinetic processes shifting from a monophase to a biphase. Measurements of fluorescence spectra showed that serine residue modification by AEBSF directly caused conspicuous changes of the tertiary structures and exposed hydrophobic surfaces. Some osmolytes were applied to find protective roles. These results confirmed that the active region of EFP is more flexible than the overall enzyme molecule and serine, as the key residue, is associated with the regional unfolding of EFP in addition to its catalytic role. The MD simulations were supportive to the kinetics data. Conclusion: Our study indicated that EFP has an essential serine residue for its catalyst function and associated folding behaviors. Also, the functional role of osmolytes such as proline and glycine that may play a role in defense mechanisms from environmental adaptation in a krill’s body was suggested. |
format |
Article in Journal/Newspaper |
author |
Qian, Guo-Ying Lim, Gyutae Yin, Shang-Jun Yang, Jun-Mo Lee, Jinhyuk Park, Yong-Doo |
spellingShingle |
Qian, Guo-Ying Lim, Gyutae Yin, Shang-Jun Yang, Jun-Mo Lee, Jinhyuk Park, Yong-Doo Biochemical Study of Fibrinolytic Protease from Euphausia superba Possessing Multifunctional Serine Protease Activity |
author_facet |
Qian, Guo-Ying Lim, Gyutae Yin, Shang-Jun Yang, Jun-Mo Lee, Jinhyuk Park, Yong-Doo |
author_sort |
Qian, Guo-Ying |
title |
Biochemical Study of Fibrinolytic Protease from Euphausia superba Possessing Multifunctional Serine Protease Activity |
title_short |
Biochemical Study of Fibrinolytic Protease from Euphausia superba Possessing Multifunctional Serine Protease Activity |
title_full |
Biochemical Study of Fibrinolytic Protease from Euphausia superba Possessing Multifunctional Serine Protease Activity |
title_fullStr |
Biochemical Study of Fibrinolytic Protease from Euphausia superba Possessing Multifunctional Serine Protease Activity |
title_full_unstemmed |
Biochemical Study of Fibrinolytic Protease from Euphausia superba Possessing Multifunctional Serine Protease Activity |
title_sort |
biochemical study of fibrinolytic protease from euphausia superba possessing multifunctional serine protease activity |
publisher |
Bentham Science Publishers Ltd. |
publishDate |
2021 |
url |
http://dx.doi.org/10.2174/0929866527666201112123714 https://eurekaselect.com/article/download/187909 https://www.eurekaselect.com/187909/article |
genre |
Euphausia superba |
genre_facet |
Euphausia superba |
op_source |
Protein & Peptide Letters volume 28, issue 6, page 651-664 ISSN 0929-8665 |
op_doi |
https://doi.org/10.2174/0929866527666201112123714 |
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Protein & Peptide Letters |
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28 |
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6 |
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651 |
op_container_end_page |
664 |
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1802643909388533760 |