Staff View: Characterization of the functional domains of FKBP35 from Plasmodium Knowlesi

Characterization of the functional domains of FKBP35 from Plasmodium Knowlesi

High incident of Plasmodium knowlesi accompanied by the increase in drug resistance cases in Malaysia Borneo urges us to develop a novel antimalarial drug with no resistance. Despite its advantages as an antimalarial compound without resistance effects, FK506 displayed an immunosuppressive side effe...

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Main Author:	Goh, Carlmond Kah Wun
Format:	Thesis
Language:	English
Published:	2019
Subjects:	QD Chemistry
Online Access:	https://eprints.ums.edu.my/id/eprint/25219/1/Characterization%20of%20the%20functional%20domains%20of%20FKBP35%20from%20Plasmodium%20Knowlesi.pdf https://eprints.ums.edu.my/id/eprint/25219/
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spelling	my.ums.eprints.252192020-03-12T08:37:21Z https://eprints.ums.edu.my/id/eprint/25219/ Characterization of the functional domains of FKBP35 from Plasmodium Knowlesi Goh, Carlmond Kah Wun QD Chemistry High incident of Plasmodium knowlesi accompanied by the increase in drug resistance cases in Malaysia Borneo urges us to develop a novel antimalarial drug with no resistance. Despite its advantages as an antimalarial compound without resistance effects, FK506 displayed an immunosuppressive side effect. In attempts to find FK506 replacers, solid fundamental studies on target molecule of FK506 are needed to provide a platform for the development of novel antimalarial drug without the risk of resistance. Previous studies revealed FK506 inhibited function of FKBP35 from P/asmodium fa/ciparum. Genome sequence of P. knowlesi suggested the presence of its FKBP35 (Pk-FKBP35) in which has high sequence similarity with FKBP35 from P. fa/ciparum (PF-FKBP35) and P. vivax (Pv-FKBP35), thus suggesting Pk-FKBP35 is considered a viable target for combating P. knowlesi. FKBP35 is a member of peptidyl-prolyl as-trans isomerase (PPiase), consisting FK506-binding domain (FKBD) followed by tetratricopeptide repeated domain (TPRD). There is no study so far on Pk-FKBP35, particularly on the functionality of these domains. In fact, most of the studies on Pf- and Pv-FKBP35 were focused on FKBD region. This is due to the finding that FKBD is the region where FK506 binds and inhibits PPiase catalytic function of this protein. Previous studies on PF-FKBP35 and Pv-FKBP35 also suggested that this protein might exhibit a dual-function of foldase and chaperonelike activities. However, involvement of the domains of FKBP35 on this dualfunction remains poorly understood. High similarity between Pk-FKBP35 and PFFKBP35 suggested that Pk-FKBP35 might also exhibit dual-function, yet no experimental evidence was reported. This study aims to understand the regulatory domains for the function of FKBP35 from Plasmodium knowlesi. To address, expression system containing gene encoding full-length of Pk-FKBP35 and its derivatives, Pk-FKBD and PK-TPRD were constructed. Each of them was expressed in Escherichia coli BL21(DE3). Two steps purifications including Ni-NTA binding affinity followed by gel filtration yielded 109, 162 and 189 mg of proteins from 1 L culture of Pk-FKBP35, Pk-FKBD, and Pk-TPRD respectively. Pk-FKBD showed comparable catalytic PPiase activities with full-length Pk-FKBP35 when tested using synthetic tetrapeptide (Suc-Ala-Leu-Pro-Phe-AMC) with kcav'KM of 4.7 x 105 M.1 s·1 2019 Thesis NonPeerReviewed text en https://eprints.ums.edu.my/id/eprint/25219/1/Characterization%20of%20the%20functional%20domains%20of%20FKBP35%20from%20Plasmodium%20Knowlesi.pdf Goh, Carlmond Kah Wun (2019) Characterization of the functional domains of FKBP35 from Plasmodium Knowlesi. Masters thesis, Universiti Malaysia Sabah.
institution	Universiti Malaysia Sabah
building	UMS Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Malaysia Sabah
content_source	UMS Institutional Repository
url_provider	http://eprints.ums.edu.my/
language	English
topic	QD Chemistry
spellingShingle	QD Chemistry Goh, Carlmond Kah Wun Characterization of the functional domains of FKBP35 from Plasmodium Knowlesi
description	High incident of Plasmodium knowlesi accompanied by the increase in drug resistance cases in Malaysia Borneo urges us to develop a novel antimalarial drug with no resistance. Despite its advantages as an antimalarial compound without resistance effects, FK506 displayed an immunosuppressive side effect. In attempts to find FK506 replacers, solid fundamental studies on target molecule of FK506 are needed to provide a platform for the development of novel antimalarial drug without the risk of resistance. Previous studies revealed FK506 inhibited function of FKBP35 from P/asmodium fa/ciparum. Genome sequence of P. knowlesi suggested the presence of its FKBP35 (Pk-FKBP35) in which has high sequence similarity with FKBP35 from P. fa/ciparum (PF-FKBP35) and P. vivax (Pv-FKBP35), thus suggesting Pk-FKBP35 is considered a viable target for combating P. knowlesi. FKBP35 is a member of peptidyl-prolyl as-trans isomerase (PPiase), consisting FK506-binding domain (FKBD) followed by tetratricopeptide repeated domain (TPRD). There is no study so far on Pk-FKBP35, particularly on the functionality of these domains. In fact, most of the studies on Pf- and Pv-FKBP35 were focused on FKBD region. This is due to the finding that FKBD is the region where FK506 binds and inhibits PPiase catalytic function of this protein. Previous studies on PF-FKBP35 and Pv-FKBP35 also suggested that this protein might exhibit a dual-function of foldase and chaperonelike activities. However, involvement of the domains of FKBP35 on this dualfunction remains poorly understood. High similarity between Pk-FKBP35 and PFFKBP35 suggested that Pk-FKBP35 might also exhibit dual-function, yet no experimental evidence was reported. This study aims to understand the regulatory domains for the function of FKBP35 from Plasmodium knowlesi. To address, expression system containing gene encoding full-length of Pk-FKBP35 and its derivatives, Pk-FKBD and PK-TPRD were constructed. Each of them was expressed in Escherichia coli BL21(DE3). Two steps purifications including Ni-NTA binding affinity followed by gel filtration yielded 109, 162 and 189 mg of proteins from 1 L culture of Pk-FKBP35, Pk-FKBD, and Pk-TPRD respectively. Pk-FKBD showed comparable catalytic PPiase activities with full-length Pk-FKBP35 when tested using synthetic tetrapeptide (Suc-Ala-Leu-Pro-Phe-AMC) with kcav'KM of 4.7 x 105 M.1 s·1
format	Thesis
author	Goh, Carlmond Kah Wun
author_facet	Goh, Carlmond Kah Wun
author_sort	Goh, Carlmond Kah Wun
title	Characterization of the functional domains of FKBP35 from Plasmodium Knowlesi
title_short	Characterization of the functional domains of FKBP35 from Plasmodium Knowlesi
title_full	Characterization of the functional domains of FKBP35 from Plasmodium Knowlesi
title_fullStr	Characterization of the functional domains of FKBP35 from Plasmodium Knowlesi
title_full_unstemmed	Characterization of the functional domains of FKBP35 from Plasmodium Knowlesi
title_sort	characterization of the functional domains of fkbp35 from plasmodium knowlesi
publishDate	2019
url	https://eprints.ums.edu.my/id/eprint/25219/1/Characterization%20of%20the%20functional%20domains%20of%20FKBP35%20from%20Plasmodium%20Knowlesi.pdf https://eprints.ums.edu.my/id/eprint/25219/
_version_	1760230339872030720
score	13.211869

Characterization of the functional domains of FKBP35 from Plasmodium Knowlesi

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