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Identification and molecular characterization of Parkin in Clonorchis sinensis.

Bai X, Kim TI, Lee JY, Dai F, Hong SJ - Korean J. Parasitol. (2015)

Bottom Line: CsParkin was localized in the locomotive and male reproductive organs of C. sinensis adults, and extensively in metacercariae.Parkin has been found to participate in regulating mitochondrial function and energy metabolism in mammalian cells.From these results, it is suggested that CsParkin play roles in energy metabolism of the locomotive organs, and possibly in protein metabolism of the reproductive organs of C. sinensis.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Environmental Biology, Chung-Ang University College of Medicine, Seoul 156-756, Korea.

ABSTRACT
Clonorchis sinensis habitating in the bile duct of mammals causes clonorchiasis endemic in East Asian countries. Parkin is a RING-between-RING protein and has E3-ubiquitin ligase activity catalyzing ubiquitination and degradation of substrate proteins. A cDNA clone of C. sinensis was predicted to encode a polypeptide homologous to parkin (CsParkin) including 5 domains (Ubl, RING0, RING1, IBR, and RING2). The cysteine and histidine residues binding to Zn(2+) were all conserved and participated in formation of tertiary structural RINGs. Conserved residues were also an E2-binding site in RING1 domain and a catalytic cysteine residue in the RING2 domain. Native CsParkin was determined to have an estimated molecular weight of 45.7 kDa from C. sinensis adults by immunoblotting. CsParkin revealed E3-ubiquitin ligase activity and higher expression in metacercariae than in adults. CsParkin was localized in the locomotive and male reproductive organs of C. sinensis adults, and extensively in metacercariae. Parkin has been found to participate in regulating mitochondrial function and energy metabolism in mammalian cells. From these results, it is suggested that CsParkin play roles in energy metabolism of the locomotive organs, and possibly in protein metabolism of the reproductive organs of C. sinensis.

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A putative tertiary structure of CsParkin. The tertiary structure of CsParkin was predicted with Phyre2 using rat parkin as a template. Conformations of Ubl, RING0, RING1, IBR, and RING2 domain are displayed in different colors. A grey sphere denotes the bound Zn2+ ion. An E2-binding region in RING1 domain is marked by a broken purple circle. REP fragment containing a key residue Trp339 is identified between IBR and RING2. The catalytic residue Cys375 is depicted with a yellow sphere.
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f2-kjp-53-1-65: A putative tertiary structure of CsParkin. The tertiary structure of CsParkin was predicted with Phyre2 using rat parkin as a template. Conformations of Ubl, RING0, RING1, IBR, and RING2 domain are displayed in different colors. A grey sphere denotes the bound Zn2+ ion. An E2-binding region in RING1 domain is marked by a broken purple circle. REP fragment containing a key residue Trp339 is identified between IBR and RING2. The catalytic residue Cys375 is depicted with a yellow sphere.

Mentions: The CsParkin cDNA was 1,218 nucleotides long and encoded a putative polypeptide of 405 amino acids with an estimated molecular weight of 45.7 kDa. In the polypeptide, highly conserved were Cys and His residues which bind to Zn2+ ion and form RING domains (Fig. 1). Secondary structure of CsParkin revealed 5 domains in tandem: ubiquitin-like (Ubl), RING0, RING1, in-between-RING (IBR), and RING2. CsParkin lacked of a long tether between the Ubl and RING0 domains, but found in other animals. In Ubl domain, a hydrophobic RING1-binding site was conserved at Leu43. RING1 was a canonical RING finger with classic cross-brace arrangement and had an E2-binding site consisting of Ala164, Phe166, and Asn167. In all parkins, a linker between IBR and RING2, termed a repressor element of parkin (REP) [26], displayed low similarity, but a residue Trp339 was highly conserved. A catalytic motif consisting of Cys375, X376, and His377 was well conserved and evident in CsParkin RING2 domain (Fig. 2).


Identification and molecular characterization of Parkin in Clonorchis sinensis.

Bai X, Kim TI, Lee JY, Dai F, Hong SJ - Korean J. Parasitol. (2015)

A putative tertiary structure of CsParkin. The tertiary structure of CsParkin was predicted with Phyre2 using rat parkin as a template. Conformations of Ubl, RING0, RING1, IBR, and RING2 domain are displayed in different colors. A grey sphere denotes the bound Zn2+ ion. An E2-binding region in RING1 domain is marked by a broken purple circle. REP fragment containing a key residue Trp339 is identified between IBR and RING2. The catalytic residue Cys375 is depicted with a yellow sphere.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4384794&req=5

f2-kjp-53-1-65: A putative tertiary structure of CsParkin. The tertiary structure of CsParkin was predicted with Phyre2 using rat parkin as a template. Conformations of Ubl, RING0, RING1, IBR, and RING2 domain are displayed in different colors. A grey sphere denotes the bound Zn2+ ion. An E2-binding region in RING1 domain is marked by a broken purple circle. REP fragment containing a key residue Trp339 is identified between IBR and RING2. The catalytic residue Cys375 is depicted with a yellow sphere.
Mentions: The CsParkin cDNA was 1,218 nucleotides long and encoded a putative polypeptide of 405 amino acids with an estimated molecular weight of 45.7 kDa. In the polypeptide, highly conserved were Cys and His residues which bind to Zn2+ ion and form RING domains (Fig. 1). Secondary structure of CsParkin revealed 5 domains in tandem: ubiquitin-like (Ubl), RING0, RING1, in-between-RING (IBR), and RING2. CsParkin lacked of a long tether between the Ubl and RING0 domains, but found in other animals. In Ubl domain, a hydrophobic RING1-binding site was conserved at Leu43. RING1 was a canonical RING finger with classic cross-brace arrangement and had an E2-binding site consisting of Ala164, Phe166, and Asn167. In all parkins, a linker between IBR and RING2, termed a repressor element of parkin (REP) [26], displayed low similarity, but a residue Trp339 was highly conserved. A catalytic motif consisting of Cys375, X376, and His377 was well conserved and evident in CsParkin RING2 domain (Fig. 2).

Bottom Line: CsParkin was localized in the locomotive and male reproductive organs of C. sinensis adults, and extensively in metacercariae.Parkin has been found to participate in regulating mitochondrial function and energy metabolism in mammalian cells.From these results, it is suggested that CsParkin play roles in energy metabolism of the locomotive organs, and possibly in protein metabolism of the reproductive organs of C. sinensis.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Environmental Biology, Chung-Ang University College of Medicine, Seoul 156-756, Korea.

ABSTRACT
Clonorchis sinensis habitating in the bile duct of mammals causes clonorchiasis endemic in East Asian countries. Parkin is a RING-between-RING protein and has E3-ubiquitin ligase activity catalyzing ubiquitination and degradation of substrate proteins. A cDNA clone of C. sinensis was predicted to encode a polypeptide homologous to parkin (CsParkin) including 5 domains (Ubl, RING0, RING1, IBR, and RING2). The cysteine and histidine residues binding to Zn(2+) were all conserved and participated in formation of tertiary structural RINGs. Conserved residues were also an E2-binding site in RING1 domain and a catalytic cysteine residue in the RING2 domain. Native CsParkin was determined to have an estimated molecular weight of 45.7 kDa from C. sinensis adults by immunoblotting. CsParkin revealed E3-ubiquitin ligase activity and higher expression in metacercariae than in adults. CsParkin was localized in the locomotive and male reproductive organs of C. sinensis adults, and extensively in metacercariae. Parkin has been found to participate in regulating mitochondrial function and energy metabolism in mammalian cells. From these results, it is suggested that CsParkin play roles in energy metabolism of the locomotive organs, and possibly in protein metabolism of the reproductive organs of C. sinensis.

Show MeSH
Related in: MedlinePlus