FcγRⅡb is the only inhibitory receptor of the IgG Fc receptor(FcγR)and plays an important role in the negative regulation and immune responses.In order to screen the binding peptide of sFcγRⅡb, the recombinant sFcγRⅡb protein was used as the target molecule, and the sFcγRⅡb binding peptide was screened by phage peptide library display technology.The affinity of each eluted phage to sFcγRⅡb protein was identified by ELISA.40 phage clones were screened after four rounds screening, and 28 different 12-residue peptides were obtained through translation of DNA sequencing data.The affinities between phages and sFcγRⅡb protein were estimated and confirmed by ELISA analysis, and the results showed that the deduced peptide sequence, FHKMPWYMSMYY was the highest specificity and affinity to sFcγRⅡb protein. Furthermore, this study may lay a foundation for the further research on the mechanism of FcγRⅡb and the function of binding peptide.
XU Zhenzhen
,
LIU Lipeng
,
ZHANG Yanfen
,
CUI Zhe
,
BI Kewei
,
LIU Zhongcheng
. Screening and identification of sFcγRⅡb binding peptide[J]. Journal of Hebei University(Natural Science Edition), 2019
, 39(4)
: 398
-403
.
DOI: 10.3969/j.issn.1000-1565.2019.04.011
[1] HOGARTH P M, PIETERSZ G A.Fc receptor-targeted therapies for the treatment of inflammation, cancer and beyond[J].Nature Reviews Drug Discovery, 2012, 11(4):311-331.DOI:10.1038/nrd2909.
[2] STARBECKMILER G R, BADOVINAC V P, BARBER D L, et al.Expression of FcγRⅡb Tempers Memory CD8 T-cell Function in vivo[J].Journal of Immunology, 2013, 192(1):35-39.DOI:10.4049/jimmunol.1302232.
[3] BOURNAZOS S, RAVETCH J V.Fcγ receptor function and the design of vaccination strategies[J].Immunity, 2017, 47(2):224-233.DOI:10.1016/j.immuni.2017.07.009.
[4] ROGHANIAN A, STOPFORTH R J, DAHAL L N, et al.New revelations from an old receptor:Immunoregulatory functions of the inhibitory Fc gamma receptor, FcγRⅡb(CD32B)[J].Journal of Leukocyte Biology, 2018, 103(6):1077-1088.DOI:10.1002/jlb.2mir0917-354r.
[5] MALBEC O, CASSARD L, ALBANESI M, et al.Trans-inhibition of activation and proliferation signals by Fc receptors in mast cells and basophils[J].Science Signaling, 2016, 9(459):ra126.DOI:10.1126/scisignal.aag1401.
[6] MALBEC O, ATTAL J P, FRIDMAN W H, et al.Negative regulation of mast cell proliferation by FcγRⅡb[J].Molecular immunology, 2002, 38(16/17/18):1295-1299.DOI:10.1016/s0161-5890(02)00078-0.
[7] BAO D K, WANG R, QIAO S L, et al.Antibody-dependent enhancement of PRRSV infection down-modulates TNF-α and IFN-β transcription in macrophages[J].Veterinary Immunology and Immunopathology, 2013, 156(1/2):128-134.DOI:10.1016/j.vetimm.2013.09.006.
[8] ESPELI M, SMITH K G C, CLATWORTHY M R.FcγRⅡb and autoimmunity[J].Immunological Reviews, 2016, 269(1):194-211.DOI:10.1111/imr.12368.
[9] SONDERMANN P.The FcγR/IgG interaction as target for the treatment of autoimmune diseases[J].Journal of Clinical Immunology, 2016, 36(1):95-99.DOI:10.1007/s10875-016-0272-7.
[10] HU X P, WU J Q, ZHU L P, et al.Association of Fcγ receptor IIB polymorphism with cryptococcal meningitis in HIV-uninfected Chinese patients[J].PLoS One, 2012, 7(8):e42439.DOI:10.1371/journal.pone.0042439.
[11] KAY B K, KASANOV J, YAMABHAI M.Screening phage-displayed combinatorial peptide libraries[J].Methods, 2001, 24(3):240-246.DOI:10.1006/meth.2001.1185.
[12] LOPES R S, QUEIROZ M A F, GOMES S T M, et al.Phage display:an important tool in the discovery of peptides with anti-HIV activity[J].Biotechnology Advances, 2018,36(7):1847-1854.DOI:10.1016/j.biotechadv.2018.07.003.
[13] LISI S, SISTO M, LOFRUMENTO D D, et al.Advances in the understanding of the Fc gamma receptors-mediated autoantibodies uptake[J].Clinical and Experimental Medicine, 2011, 11(1):1-10.DOI:10.1007/s10238-010-0098-1.
[14] 张雷, 曹秀琴, 杨志伟.人可溶型FcγRⅡb的制备及其功能初步研究[J].细胞与分子免疫学杂志, 2012, 28(9):952-955.DOI:10.13423/j.cnki.cjcmi.006547.
[15] 曹虹, 林琳, 刘北一,等.脑膜炎大肠杆菌IbeA蛋白结合肽序列的亲和筛选[J].微生物学通报, 2006, 33(5):101-106.DOI:10.3969/j.issn.0253-2654.2006.05.021.
[16] MEULEMANS E V, SLOBBE R, WASTERVAL P, et al.Selection of phage-displayed antibodies specific for a cytoskeletal antigen by competitive elution with a monoclonal antibody[J].Journal of Molecular Biology, 1994, 244(4):353-360.DOI:10.1006/jmbi.1994.1735.
[17] 王祥卫, 郭应禄, 张立新, 等.人膀胱癌P-糖蛋白结合肽的筛选[J].中华泌尿外科杂志, 2003, 24(12):824-826.DOI:10.3760/j:issn:1000-6702.2003.12.010.
[18] ZHANG F, WANG M, QIU Z, et al.Identification and characterization of strychnine-binding peptides using phage-display screening[J].Protein & Peptide Letters, 2017, 24(7):626-632.DOI:10.2174/0929866524666170404164408.