title: Expression analysis and enzymatic characterization of phospholipase Cδ4 from olive flounder (Paralichthys olivaceus).
abstract: Phospholipase Cδ4 (PLCδ4) plays a significant role in cell proliferation, tumorigenesis, and in an early stage of fertilization. Despite the characterization of the mammalian PLCδ4, extensive study in aquatic organisms has not been carried out so far. Here, we performed the molecular and biochemical characterization of flatfish Paralichthys olivaceus PLCδ4 (PoPLCδ4) to understand its enzymatic properties and physiological functions. The olive flounder PLCδ4 cDNA has an open reading frame (ORF) of 2,268 bp, and encodes a 755 amino acid polypeptide with a predicted molecular weight of 86 kDa. All the characteristic domains found in mammalian PLCδ isoforms (PH domain, EF hands, an X-Y catalytic region, and a C2 domain) were found to be present in PoPLCδ4. The mRNA expression analysis of PoPLCδ4 showed that PoPLCδ4 is predominantly expressed in the brain, eye and heart tissues. Like other mammalian PLCδ proteins, the enzyme activity of recombinant PoPLCδ4 to phosphatidylinositol-4,5-bis-phosphate (PIP2) was noted to be concentration- and Ca(2+)-dependent. The structural features and biochemical characteristics of PoPLCδ4 were found to be similar to those of mammalian PLCδ4. This is the first demonstration of the expression analysis and enzymatic characterization of piscine PLCδ4.
authors.affiliation: Department of Biotechnology, Pukyong National University, Busan 608-737, South Korea.
authors.forename: Hye Jin
authors.initials: HJ
authors.lastname: Bak
authors.forename: Moo-Sang
authors.initials: MS
authors.lastname: Kim
authors.forename: Na Young
authors.initials: NY
authors.lastname: Kim
authors.forename: A Ram
authors.initials: AR
authors.lastname: Lee
authors.forename: Ju Hyeon
authors.initials: JH
authors.lastname: Park
authors.forename: Jin Young
authors.initials: JY
authors.lastname: Lee
authors.forename: Bo Seong
authors.initials: BS
authors.lastname: Kim
authors.forename: Sang Jung
authors.initials: SJ
authors.lastname: Ahn
authors.forename: Hyung Ho
authors.initials: HH
authors.lastname: Lee
authors.forename: Joon Ki
authors.initials: JK
authors.lastname: Chung
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D005412:Flatfishes / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic; D008969:Molecular Sequence Data; D009928:Organ Specificity; D054803:Phospholipase C delta / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D021381:Protein Transport; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D011994:Recombinant Proteins; D054803:Phospholipase C delta
keywords: Immunohistochemistry; Olive flounder (Paralichthys olivaceus); PLCδ4; Phosphatidylinositol-4,5-bis-phosphate (PIP(2)); mRNA expression