鸡γ-干扰素与补体受体2蛋白相互作用的研究

doi:10.16431/j.cnki.1671-7236.2024.10.028

Abstract

Abstract: 【Objective】 The experiment was to study the interaction between chicken interferon γ (chIFN-γ) and chicken complement receptor 2 (chCR2) proteins，and lay a foundation for the research of new functions of chIFN-γ and chCR2.【Method】 chIFN-γ related expression plasmid was constructed by homologous recombination method. The interaction between chIFN-γ and chCR2 proteins was identified by co-immunoprecipitation，laser confocal and surface plasmon resonance techniques，and the interaction mode of chIFN-γ and chCR2 proteins was simulated by molecular docking.【Result】 The pCMV-Myc-chIFN-γ expression plasmid was constructed successfully. The plasmid of pCMV-Myc-chIFN-γ and pCMV-HA-chCR2-ΔTM were co-transfected into HEK-293FT cells and the results showed that chIFN-γ could interact with chCR2. The plasmid of pCMV-Myc-chIFN-γ and pCMV-HA-chCR2-ΔTM were co-transfected into DF-1 cells and the results showed that chIFN-γ and chCR2 proteins could co-locate in DF-1 cells. The results of surface plasmon resonance test showed that the equilibrium dissociation constant (KD) between chIFN-γ and chCR2 proteins was 0.362 μmol/L，and the affinity between the two proteins was high. The results of molecular docking experiments showed that the interaction between chIFN-γ and chCR2 proteins was 1 salt bridge between Asp23-His150 and 3 sets of hydrogen bonds between Asn37-Ser143，Glu57-Gly179 and Ile93-Tyr184.【Conclusion】 The results of this study proved that chIFN-γ and chCR2 proteins could interact，and the interaction mode between them was 1 salt bridge and 3 sets of hydrogen bonds.

Key words: chicken interferon γ (chIFN-γ); chicken complement receptor 2 (chCR2); interaction; affinity

CLC Number:

S831

IN Huan, MENG Zhaoying, TU Min, SHEN Jia, SHI Aihua, ZHAO Lei, XI Shuo, ZHANGJianwei, ZHANG Zhenhua. Study on the Interaction Between Chicken Interferon γ and Complement Receptor 2 Protein[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(10): 4476-4484.

References

[1] LUKACSI S，MACSIK V B，NAGY B Z，et al. Utilization of complement receptors in immune cell-microbe interaction[J]. FEBS Letters,2020,594(16):2695-2713.
[2] HANNAN J P. The structure-function relationships of complement receptor type 2(CR2; CD21)[J]. Current Protein & Peptide Science,2016,17: 463-487.
[3] TEW J G，WU J，QIN D，et al. Follicular dendritic cells and presentation of antigen and costimulatory signals to B cells[J]. Immunological Reviews,1997,156:39-52.
[4] NIELSEN C H，FISCHER E M，LESLIE R G. The role of complement in the acquired immune response[J]. Immunology,2000,100(1):4-12.
[5] YOUNG K A，CHEN X S，HOLERS V M，et al. Isolating the Epstein-Barr virus gp350/220 binding site on complement receptor type 2(CR2/CD21)[J]. Journal of Biological Chemistry，2007,282(50):36614-36625.
[6] PESTKA S，LANGER J A，ZOON K C，et al. Interferons and their actions[J]. Annual Review of Biochemistry,1987,56: 727-777.
[7] BILLIAU A，MATTHYS P. Interferon-gamma: A historical perspective[J].Cytokine & Growth Factor Reviews,2009,20:97-113.
[8] CAPOBIANCHI M R，ULERI E，CAGLIOTI C，et al. Type Ⅰ IFN family members: Similarity，differences and interaction[J]. Cytokine & Growth Factor Reviews,2015,26(2):103-111.
[9] BROGGI A，GRANUCCI F，ZANONI I. Type Ⅲ interferons: Balancing tissue tolerance and resistance to pathogen invasion[J].Journal of Experimental Medicine,2020,217(1): e20190295.
[10] LIU W，ZHANG S，WANG J. IFN-γ，should not be ignored in SLE[J]. Frontiers in Immunology,2022,13:954706.
[11] POGUE S L，PRESTON B T，STALDER J，et al. The receptor for type Ⅰ IFNs is highly expressed on peripheral blood B cells and monocytes and mediates a distinct profile of differentiation and activation of these cells[J].Journal of Interferon & Cytokine Research,2004,24: 131-139
[12] ASOKAN R，HUA J，YOUNG K A，et al. Characterization of human complement receptor type 2(CR2/CD21) as a receptor for IFN-alpha: A potential role in systemic lupus erythematosus[J].The Journal of Immunology, 2006,177(1):383-394.
[13] SUGANO N，CHEN W，ROBERTS M L,et al. Cooper. Epstein-Barr virus binding to CD21 activates the initial viral promoter via NF-κB induction[J]. Journal of Experimental Medicine,1997,186: 731-737.
[14] ADDARIO M，LIBERMANN T A，XU J，et al. Epstein-Barr virus and its glycoprotein-350 upregulate IL-6 in human B-lymphocytes via CD21，involving activation of NF-κB and different signaling pathways[J].Journal of Molecular Biology，2001,308: 501-514.
[15] AUBRY J P，POCHON S，GRABER P，et al. CD21 is a ligand for CD23 and regulates IgE production[J]. Nature,1992,358: 505-507.
[16] DELCAYRE A X，SALAS F，MATHUR S，et al. Epstein-Barr virus/complement C3d receptor is an interferon alpha receptor[J]. The EMBO Journal,1991,10: 919-926.
[17] DARNELL J E，KERR I M，STARK G R. Jak-STAT pathways and transcriptional activation in response to IFNs and other extracellular signaling proteins[J].Science,1994,264:1415-1421.
[18] PINI L，RASTELLI G. Molecular docking: Shifting paradigms in drug discovery[J]. International Journal of Molecular Sciences,2019,20(18):4331.
[19] FENG Y，YAN Y，HE J，et al. Docking and scoring for nucleic acid-ligand interactions: Principles and current status[J]. Drug Discovery Today,2022,27(3):838-847.
[20] FENG Y，ZHANG K，WU Q，et al. NLDock: A fast nucleic acid-ligand docking algorithm for modeling RNA/DNA-ligand complexes[J].Journal of Chemical Information and Modeling,2021,61(9):4771-4782.
[21] IIDA K，NADLER L，NUSSENZWEIG V. Identification of the membrane receptor for the complement fragment C3d by means of a monoclonal antibody[J].Journal of Experimental Medicine,1983,158:1021-1033.
[22] FINGEROTH J D，WEIS J J，TEDDER T F，et al. Epstein-Barr virus receptor of human B lymphocytes is the C3d receptor CR2[J]. Proceedings of the National Academy of Sciences of the United States of America,1984,81:4510-4514.
[23] AUBRY J P，POCHON S，GAUCHAT J F，et al. CD23 interacts with a new functional extracytoplasmic domain involving N-linked oligosaccharides on CD21[J]. Journal of Immunology,1994,152: 5806-5813.
[24] JIN H，KONG Z，JIANG B，et al. Identification and characterization of chCR2，a protein that binds chicken complement component 3d[J].Journal of Immunology，2023,210(9):1408-1418.
[25] RAUW F，GARDIN Y，PALYA V，et al. The positive adjuvant effect of chitosan on antigen-specific cell-mediated immunity after chickens vaccination with live Newcastle disease vaccine[J]. Veterinary Immunology and Immunopathology,2010,134(3-4):249-258.
[26] FERREIRA L G，DOS SANTOS R N，OLIYA G，et al. Molecular docking and structure-based drug design strategies[J]. Molecules,2015,20(7):13384-13421.
[27] FERREIA L G，DOS SANTOS R N，OLIVA G，et al. Molecular docking and structure-based drug design strategies[J].Molecules,2015,20(7):13384-13421.
[28] MARRAKCHI H，LANEELLE G, QUEMARD A. InhA，a target of the antituberculous drug isoniazid，is involved in a mycobacterial fatty acid elongation system，FAS-Ⅱ[J].Microbiology,2000,146:289-296.
[29] WAHID A A，DUNPHY R W，MACPHERSON A，et al. Insights into the structure-function relationships of dimeric C3d fragments[J].Frontiers in Immunology,2021,12:714055.
[30] JIN H，TU M，MENG Z，et al. Identification and structural analysis of dimeric chicken complement component 3d and its binding with chicken complement receptor 2[J].Developmental & Comparative Immunology,2024,152:105109.
[31] TOAPANTA F R，ROSS T M. Complement-mediated activation of the adaptive immune responses: Role of C3d in linking the innate and adaptive immunity[J].Immunologic Research,2006,36(1-3)：197-210.
[32] LI K，OKEMEFUNA A I，GOR J，et al.Solution structure of the complex formed between human complement C3d and full-length complement receptor type 2[J]. Journal of Molecular Biology,2008,384(1)：137-150.
[33] CARTER R H，FEARON D T. Polymeric C3dg primes human B lymphocytes for proliferation induced by anti-IgM[J].The Journal Immunology,1989,143(6)：1755-1760.
[34] BOWER J F,YANG X，SODROSKI J,et al. Elicitation of neutralizing antibodies with DNA vaccines expressing soluble stabilized Human immunodeficiency virus type 1 envelope glycoprotein trimers conjugated to C3d[J].Journal of Virology,2004,78(9)：4710-4719.
[35] GREEN T D，NEWTON B R，ROTA P A，et al. C3d enhancement of neutralizing antibodies to measles hemagglutinin[J].Vaccine,2001,20(1-2)：242-248.
[36] ZHANG Z，LI Y，XU S，et al. Fusion to chicken C3d enhances the immunogenicity of the M2 protein of Avian influenza virus[J].Journal of Virology,2010,7:89.
[37] 都兴洋. 表达鸡IFN-γ基因重组鸡痘病毒与鸡传染性法氏囊疫苗联合接种对鸡免疫应答的影响[D].呼和浩特：内蒙古农业大学,2006.DU X Y. Effect of rFPV-IFN-γ on active immunization induced by infectious bursal disease live vaxxine in chickens[D].Hohhot:Inner Mongolia Agricultural University,2006.(in Chinese)

Study on the Interaction Between Chicken Interferon γ and Complement Receptor 2 Protein

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	CUI Jinqiang, CHENG Jing, JIANG Bo, ZHOU Linyi, LIU Wenxiao, LI Yongqing, SONG Qinye. Prokaryotic Expression of Chicken Interferon-γ and Preparation and Identification of Its Monoclonal Antibodies [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(7): 3008-3019.
[2]	LI Siyao, HOU Bin, MALI Yaqiqige, HASI Surong. Research Progress on the Interaction Between Intestinal Parasites and Intestinal Microorganisms [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(1): 330-337.
[3]	YU Xin, HU Yumiao, LI Fangbing, YANG Yadong, ZHAO Jin, FENG Yali, ZHANG Ying. Cloning and Bioinformatics Analysis of TLR4 Gene in Cygnus olor [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(9): 3469-3479.
[4]	LI Mengrui, LI Yongxia, LIN Xiaofeng, HUANG Xiaohang, HUANG Zhijian, YIN Guangwen. Research Progress on the Interaction Mechanism Between Parasites and Gut Microbiota [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1129-1139.
[5]	LIANG Shasha, YU Nongqi, LU Ying, LI Tingting, YAN Shengfei, HUANG Jian, TAN Zhengzhun, HUANG Rongchun, LI Hui, PAN Weijun, QIN Guangsheng. Screening of Key Genes Related to Intramuscular Fat Deposition in Buffaloes Based on GEO Database and Bioinformatics Analysis [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4359-4369.
[6]	LUO Lan, CUI Chao, DONG Ruilan, LIU Haoyu, LIU Xiaobin, YU Guanghui. Effects of Different Concentrations of Rutin and Freezing Rates on Cryopreservation of Boar Semen [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 982-991.
[7]	ZHANG Yiqing, LI Juan, QI Nanshan, LIAO Shenquan, LYU Minna, LIN Xuhui, CAI Haiming, HU Junjing, ZHANG Jianfei, LI Yugu, SUN Mingfei. Mechanism of lncRNA in Apicomplexan Parasites Infection [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4392-4400.
[8]	LIU Zaixia, DUAN Shi, SUN Yanyong, LYU Qi, FU Shaoyin, HE Xiaolong, ZHANG Wenguang, LIU Yongbin. Study on Differential Gene and Protein Interaction Network of Testis in Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(1): 1-11.
[9]	SUN Aoying, WANG Yanan, JIANG Yanping, CUI Wen, QIAO Xinyuan, ZHOU Han, TANG Lijie, XU Yigang, LI Yijing, WANG Li. Study on Interaction Between Transmissible Gastroenteritis Virus Nsp2 and Host Cell Protein PSMD11 [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(7): 2569-2576.
[10]	HAN Fei, ZHAO Ruixiao, WANG Gang, JIANG Mingfeng. Prokaryotic Expression and Identification of Porcine Derived Glucagon Like Peptide-2 Fusion Protein with Site Directed Mutagenesis [J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(11): 3137-3143.
[11]	ZHOU Yongchun, WANG Kunlun, YAN Yaqun, ZHAO Shanshan, NING Changshen. Advances on the Interaction Between Intestinal Parasites and Flora [J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(10): 2876-2881.
[12]	WU Suiqin, HUANG Shan, GAO Cheng, MENG Hui. Study on Rapid Detection of Chloramphenicol by sol-gel Column in Shrimp [J]. , 2018, 45(10): 2894-2901.
[13]	BAI Dongdong, LI Xinpu, YANG Feng, LUO Jinyin, WANG Xurong, LI Hongsheng. The Mechanism Analysis of Baitouweng Decoction in Treating Swine Diarrhea Based on Network Pharmacology [J]. , 2018, 45(10): 2866-2875.
[14]	HUANG Kongwei, LI Zhipeng, CUI Kuiqing, SHI Deshun, LIU Qingyou. Prokaryotic Expression and Purification of Recombinant Hirudin [J]. , 2018, 45(1): 1-13.
[15]	ZHOU Chong, HUANG Li, SHI De-shun, JIANG Jian-rong. Effects of Mifepristone (RU486) on Sperm/Cumulus Penetration in Mice [J]. , 2017, 44(5): 1275-1280.