title: Myostatin inhibitory region of fish (Paralichthys olivaceus) myostatin-1 propeptide.
abstract: Myostatin (MSTN) is a potent negative regulator of skeletal muscle growth, and its activity is suppressed by MSTN propeptide (MSTNpro), the N-terminal part of MSTN precursor cleaved during post-translational MSTN processing. The current study examined which region of flatfish (Paralichthys olivaceus) MSTN-1 propeptide (MSTN1pro) is critical for MSTN inhibition. Six different truncated forms of MSTN1pro containing N-terminal maltose binding protein (MBP) as a fusion partner were expressed in Escherichia coli, and partially purified by an affinity chromatography for MSTN-inhibitory activity examination. Peptides covering different regions of flatfish MSTN1pro were also synthesized for MSTN-inhibitory activity examination. A MBP-fused MSTN1pro region consisting of residues 45-100 had the same MSTN-inhibitory potency as the full sequence flatfish MSTN1pro (residues 23-265), indicating that the region of flatfish MSTN1pro consisting of residues 45-100 is sufficient to maintain the full MSTN-inhibitory capacity. A MBP-fused MSTN1pro region consisting of residues 45-80 (Pro45-80) also showed MSTN-inhibitory activity with a lower potency, and the Pro45-80 demonstrated its MSTN binding capacity in a pull-down assay, indicating that the MSTN-inhibitory capacity of Pro45-80 is due to its binding to MSTN. Flatfish MSTN1pro synthetic peptides covering residues 45-65, 45-70, and 45-80 demonstrated MSTN-inhibitory activities, but not the synthetic peptide covering residues 45-54, indicating that residues 45-65 of flatfish MSTN1pro are essential for MSTN inhibition. In conclusion, current study show that like the mammalian MSTNpro, the MSTN-inhibitory region of flatfish MSTN1pro resides near its N-terminus, and imply that smaller sizes of MSTNpro can be effectively used in various applications designed for MSTN inhibition.
authors.affiliation: Department of Marine Molecular Biotechnology, Gangneung-Wonju National University, Gangneung-si, Ganwon-do, 210-702, South Korea; Department of Human Nutrition, Food and Animal Sciences, University of Hawaii, 1955 East-West Rd., Honolulu, HI 96822, USA.
authors.forename: Sang Beum
authors.initials: SB
authors.lastname: Lee
authors.affiliation: Department of Marine Molecular Biotechnology, Gangneung-Wonju National University, Gangneung-si, Ganwon-do, 210-702, South Korea.
authors.forename: Jeong Hwan
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Department of Marine Molecular Biotechnology, Gangneung-Wonju National University, Gangneung-si, Ganwon-do, 210-702, South Korea.
authors.forename: Deuk-Hee
authors.initials: DH
authors.lastname: Jin
authors.affiliation: Department of Marine Molecular Biotechnology, Gangneung-Wonju National University, Gangneung-si, Ganwon-do, 210-702, South Korea. Electronic address: hj-jin@gwnu.ac.kr.
authors.forename: Hyung-Joo
authors.initials: HJ
authors.lastname: Jin
authors.affiliation: Department of Human Nutrition, Food and Animal Sciences, University of Hawaii, 1955 East-West Rd., Honolulu, HI 96822, USA. Electronic address: ykim@hawaii.edu.
authors.forename: Yong Soo
authors.initials: YS
authors.lastname: Kim
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D029941:Fish Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005412:Flatfishes / Q000378:metabolism*; D017930:Genes, Reporter / Q000187:drug effects; D057809:HEK293 Cells; D006801:Humans; D008024:Ligands; D058619:Maltose-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D008970:Molecular Weight; D055435:Myostatin / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D010455:Peptides / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D015202:Protein Engineering; D020033:Protein Isoforms / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011498:Protein Precursors / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D021382:Protein Sorting Signals / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D029941:Fish Proteins; D008024:Ligands; D058619:Maltose-Binding Proteins; D055435:Myostatin; D009842:Oligopeptides; D010446:Peptide Fragments; D010455:Peptides; D020033:Protein Isoforms; D011498:Protein Precursors; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins
keywords: Fish myostatin propeptide; Inhibitory region; Myostatin; Myostatin propeptide; Paralichthys olivaceus; Truncated protein