title: [Cloning and functional analysis of melanocortin 4 receptor mutation gene F261S].
abstract: OBJECTIVE
  To evaluate the function change of the melanocortin 4 receptor (MC4R) protein with mutation of F261S.
  
  
  METHODS
  Human embryonic cells of the HEK293 line were cultured. Wild-type genomic DNA and F261S mutation human melanocortin 4 receptor genes from the genomic DNA of aproband of homozygotic F612 mutation were amplified and cloned into a topo-TA eukaryotic expression plasmid vector. After the wild-type and F261S mutated proteins were expressed in HEK293 cells, alpha-MSH (10(-11) approximately 10(-5) mmol/L) was added, then the intracellular cAMP was detected with dual luciferase reporter assay system.
  
  
  RESULTS
  When the concentration of alpha-MSH added was 10(-9) approximately 10(-8) mmol/L, the intracellular alpha-MSH concentration of the cells transfected with wild-type MC4R gene was significantly higher than that of the cells transfected with F261S mutation gene (P < 0.05). When the concentration of alpha-MSH added went to 10(-7) approximately 10(-5) mmol/L, the differences became even more significant (all P < 0.01).
  
  
  CONCLUSIONS
  The novel MC4R mutation F261S undermines the signal transduction. It may be the possible reason leading to monogenic mutation obesity in Chinese.
authors.affiliation: Shanghai Diabetes Institute, Shanghai Jiaotong University Affiliated Sixth Hospital, Shanghai 200233, China.
authors.forename: Xin-Yu
authors.initials: XY
authors.lastname: Shao
authors.forename: Wei-Ping
authors.initials: WP
authors.lastname: Jia
authors.forename: Shu-Bing
authors.initials: SB
authors.lastname: Cai
authors.forename: Qi-Chen
authors.initials: QC
authors.lastname: Fang
authors.forename: Rong
authors.initials: R
authors.lastname: Zhang
authors.forename: Jun-Xi
authors.initials: JX
authors.lastname: Lu
authors.forename: Kun-San
authors.initials: KS
authors.lastname: Xiang
mesh_terms: D000328:Adult; D000368:Aged; D002648:Child; D002681:China; D003001:Cloning, Molecular; D005260:Female; D006801:Humans; D008297:Male; D020125:Mutation, Missense; D009765:Obesity / Q000235:genetics*; D010375:Pedigree; D044105:Receptor, Melanocortin, Type 4 / Q000235:genetics* / Q000378:metabolism / Q000502:physiology
chemical_list: C470072:MC4R protein, human; D044105:Receptor, Melanocortin, Type 4