A resistin binding peptide selected by phage display inhibits 3T3-L1 preadipocyte differentiation.
BACKGROUND
Resistin, a newly discovered cysteine-rich hormone secreted mainly by adipose tissues, has been proposed to form a biochemical link between obesity and type 2 diabetes. However, the resistin receptor has not yet been identified. This study aimed to identify resistin binding proteins/receptor.


METHODS
Three cDNA fragments with the same 11 bp 5' sequence were found by screening a cDNA phage display library of rat multiple tissues. As the reading frames of the same 11 bp 5' sequence were interrupted by a TGA stop codon, plaque lift assay was consequently used to prove the readthrough phenomenon. The stop codon in the same 11 bp 5' sequence was replaced by tryptophan, and the binding activity of the coded peptide [AWIL, which was designated as resistin binding peptide (RBP)] with resistin was identified by the confocal microscopy technique and the affinity chromatography experiment. pDual GC-resistin and pDual GC-resistin binding peptide were co-transfected into 3T3-L1 cells to confirm the function of resistin binding peptide.


RESULTS
Three cDNA fragments with the same 11 bp 5' sequence were found. The TGA stop codon in reading frames of the same 11 bp 5' sequence was proved to be readthroughed. The binding activity of RBP with resistin was consequently identified. The expression of the resistin binding peptide in 3T3-L1 preadipocytes expressing pDual GC-resistin significantly inhibited the adipogenic differentiation.


CONCLUSIONS
RBP could effectively rescue the promoted differentiation of resistin overexpressed 3T3-L1 preadipocyte.
Liu
Feng
F
Department of Pediatrics, Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China.
Guo
Xi-rong
XR
Gong
Hai-xia
HX
Ni
Yu-hui
YH
Fei
Li
L
Pan
Xiao-qin
XQ
Guo
Mei
M
Chen
Rong-hua
RH
D041721:3T3-L1 Cells; D017667:Adipocytes / Q000166:cytology / Q000187:drug effects*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects*; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D051381:Rats; D052243:Resistin / Q000037:antagonists & inhibitors / Q000378:metabolism*
D002352:Carrier Proteins; D019151:Peptide Library; D052243:Resistin; C499622:Retn protein, rat