Effects of metal ions on the structure and activity of a human anti-cyclin D1 single-chain variable fragment AD5

Yang,Ning; Yao,Nannan; Liao,Xiangzhi; Xie,Xiaona; Wu,Yan; Fan,Chuanxi; Zhao,Jialiang; Li,Guiying

doi:10.3892/mmr.2017.6756

Effects of metal ions on the structure and activity of a human anti-cyclin D1 single-chain variable fragment AD5

Authors:
- Ning Yang
- Nannan Yao
- Xiangzhi Liao
- Xiaona Xie
- Yan Wu
- Chuanxi Fan
- Jialiang Zhao
- Guiying Li
View Affiliations

Affiliations: Key Laboratory for Molecular Enzymology and Engineering of The Ministry of Education, School of Life Sciences, Jilin University, Changchun, Jilin 130012, P.R. China, Department of Endocrinology, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China
Published online on: June 12, 2017 https://doi.org/10.3892/mmr.2017.6756
Pages: 1314-1320
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Cyclin D1 has become a potential target for anti-tumor therapy. Recently, a novel human anti‑cyclin D1 single‑chain variable fragment (AD5) was identified, which demonstrated specific binding activity to cyclin D1 and exhibited anti‑tumor effects. However, the detailed characteristics of AD5 remain unclear. In the present study, the structure and activity of AD5 in the presence of copper II (Cu2+) or iron III (Fe3+) metal ions was investigated by fluorescence spectroscopy, synchronous fluorescence and enzyme‑linked immunosorbent assay. Cu2+ and Fe3+ were able to bind to AD5 and quench the fluorescence intensity of AD5 primarily by static quenching, which slightly altered the conformation of AD5 at temperatures of 293, 298 and 303 K; however, these temperatures demonstrated different effects on the activity of AD5. These results may be of value for the clinical application of anti-cyclin D1 single chain antibodies in the future.

View Figures

View References

1	Casimiro MC, Velasco-Velázquez M, Aguirre-Alvarado C and Pestell RG: Overview of cyclin D1 function in cancer and the CDK inhibitor landscape: Past and present. Expert Opin Investig Drugs. 23:295–304. 2014. View Article : Google Scholar : PubMed/NCBI
2	Choi YJ, Li X, Hydbring P, Sanda T, Stefano J, Christie AL, Signoretti S, Look AT, Kung AL, von Boehmer H and Sicinski P: The requirement for cyclin D function in tumor maintenance. Cancer Cell. 22:438–451. 2012. View Article : Google Scholar : PubMed/NCBI
3	Beroukhim R, Mermel CH, Porter D, Wei G, Raychaudhuri S, Donovan J, Barretina J, Boehm JS, Dobson J, Urashima M, et al: The landscape of somatic copy-number alteration across human cancers. Nature. 463:899–905. 2010. View Article : Google Scholar : PubMed/NCBI
4	Musgrove EA, Caldon CE, Barraclough J, Stone A and Sutherland RL: Cyclin D as a therapeutic target in cancer. Nat Rev Cancer. 11:558–572. 2011. View Article : Google Scholar : PubMed/NCBI
5	Casimiro MC, Crosariol M, Loro E, Li Z and Pestell RG: Cyclins and cell cycle control in cancer and disease. Genes Cancer. 3:649–657. 2012. View Article : Google Scholar : PubMed/NCBI
6	Roy PG, Pratt N, Purdie CA, Baker L, Ashfield A, Quinlan P and Thompson AM: High CCND1 amplification identifies a group of poor prognosis women with estrogen receptor positive breast cancer. Int J Cancer. 127:355–360. 2010.PubMed/NCBI
7	Alao JP: The regulation of cyclin D1 degradation: Roles in cancer development and the potential for therapeutic invention. Mol Cancer. 6:242007. View Article : Google Scholar : PubMed/NCBI
8	Ahmad ZA, Yeap SK, Ali AM, Ho WY, Alitheen NB and Hamid M: scFv antibody: Principles and clinical application. Clin Dev Immunol. 2012:9802502012. View Article : Google Scholar : PubMed/NCBI
9	Scott AM, Wolchok JD and Old LJ: Antibody therapy of cancer. Nat Rev Cancer. 12:278–287. 2012. View Article : Google Scholar : PubMed/NCBI
10	Elvin JG, Couston RG and van der Walle CF: Therapeutic antibodies: Market considerations, disease targets and bioprocessing. Int J Pharm. 440:83–98. 2013. View Article : Google Scholar : PubMed/NCBI
11	Holliger P and Hudson PJ: Engineered antibody fragments and the rise of single domains. Nat Biotechnol. 23:1126–1136. 2005. View Article : Google Scholar : PubMed/NCBI
12	Weisser NE and Hall JC: Applications of single-chain variable fragment antibodies in therapeutics and diagnostics. Biotechnol Adv. 27:502–520. 2009. View Article : Google Scholar : PubMed/NCBI
13	Wu Y, Zou D, Cao Y, Yao N, Wang J, Wang W, Jiang H and Li GY: Expression and purification of a human anti-cyclin D1 single-chain variable fragment antibody AD5 and its characterization. Int J Mol Med. 32:1451–1457. 2013.PubMed/NCBI
14	Zhou LH, Zhu X, Cao YH, Wang L, Chen Y, Du Br and Li GY: Construction of expression vector for anti-cyclin D1 intrabody AD5N and its inhibitory effects on cell proliferation of breast cancer. Chin J Immunol. 24:703–706. 2008.
15	Zhou LH, Zhu X, Cao YH, Chen Y, Tian Y, Wang L and Li GY: Effects of anti-cyclin D1 intrabody AD5N on HeLa cells of uterine cervix cancer. Chin J Clin Oncol. 35:942–944. 2008.
16	Avanti C, Oktaviani NA, Hinrichs WL, Frijlink HW and Mulder FA: Aspartate buffer and divalent metal ions affect oxytocin in aqueous solution and protect it from degradation. Int J Pharm. 444:139–145. 2013. View Article : Google Scholar : PubMed/NCBI
17	Tainer JA, Roberts VA and Getzoff ED: Protein metal-binding sites. Curr Opin Biotechnol. 3:378–387. 1992. View Article : Google Scholar : PubMed/NCBI
18	Trisler K, Looger LL, Sharma V, Baker M, Benson DE, Trauger S, Schultz PG and Smider VV: A metalloantibody that irreversibly binds a protein antigen. J Biol Chem. 282:26344–26353. 2007. View Article : Google Scholar : PubMed/NCBI
19	Iverson BL, Iverson SA, Roberts VA, Getzoff ED, Tainer JA, Benkovic SJ and Lerner RA: Metalloantibodies. Science. 249:659–662. 1990. View Article : Google Scholar : PubMed/NCBI
20	Zhang YP, Shi SY, Huang KL, Chen XQ and Peng MJ: Effect of Cu²⁺ and Fe³⁺ for drug delivery: Decreased binding affinity of ilaprazole to bovine serum albumin. J Lumin. 131:1927–1931. 2011. View Article : Google Scholar
21	Li GY, Zou DS and Zhou LH: Expression and purification of recombinant human cyclin D1 in E. coli BL21. J Jilin Univ. 44:839–843. 2006.
22	Lakowicz JR: Principles of Fluorescence Spectroscopy. Plenum Press; New York, NY: pp. 260–266. 1983
23	Bian H, Li M, Yu Q, Chen Z, Tian J and Liang Q: Study of the interaction of artemisinin with bovine serum albumin. Int J Biol Macromol. 39:291–297. 2006. View Article : Google Scholar : PubMed/NCBI
24	Liu Y, Chen M, Bian G, Liu J and Song L: Spectroscopic investigation of the interaction of the toxicant, 2-naphthylamine, with bovine serum albumin. J Biochem Mol Toxicol. 25:362–368. 2011. View Article : Google Scholar : PubMed/NCBI
25	Zhao X, Sheng F, Zheng J and Liu R: Composition and stability of anthocyanins from purple solanum tuberosum and their protective influence on Cr(VI) targeted to bovine serum albumin. J Agric Food Chem. 59:7902–7909. 2011. View Article : Google Scholar : PubMed/NCBI
26	Lu D, Zhao X, Zhao Y, Zhang B, Zhang B, Geng M and Liu R: Binding of Sudan II and Sudan IV to bovine serum albumin: Comparison studies. Food Chem Toxicol. 49:3158–3164. 2011. View Article : Google Scholar : PubMed/NCBI
27	Miller JN: Recent advances in molecular luminescence analysis. Proc Anal Div Chem Soc. 16:203–208. 1979.
28	Ye ZW, Ying Y, Yang XL, Zheng ZQ, Shi JN, Sun YF and Huang P: A spectroscopic study on the interaction between the anticancer drug erlotinib and human serum albumin. J Incl Phenom Macrocycl Chem. 78:405–413. 2014. View Article : Google Scholar
29	Cao X, Dong D, Liu J, Jia C, Liu W and Yang W: Studies on the interaction between triphenyltin and bovine serum albumin by fluorescence and CD spectroscopy. Chemosphere. 26–Jan;2013.[Epub ahead of Print]. View Article : Google Scholar
30	Fu M, Wang C, Li Z, Sakamaki T and Pestell RG: Minireview: Cyclin D1: Normal and abnormal functions. Endocrinology. 145:5439–5447. 2004. View Article : Google Scholar : PubMed/NCBI
31	Neumeister P, Pixley FJ, Xiong Y, Xie H, Wu K, Ashton A, Cammer M, Chan A, Symons M, Stanley ER and Pestell RG: Cyclin D1 governs adhesion and motility of macrophages. Mol Biol Cell. 14:2005–2015. 2003. View Article : Google Scholar : PubMed/NCBI
32	Malumbres M: Cell cycle-based therapies move forward. Cancer Cell. 22:419–420. 2012. View Article : Google Scholar : PubMed/NCBI
33	Ding Z, Wu CJ, Chu GC, Xiao Y, Ho D, Zhang J, Perry SR, Labrot ES, Wu X, Lis R, et al: SMAD4-dependent barrier constrains prostate cancer growth and metastatic progression. Nature. 470:269–273. 2011. View Article : Google Scholar : PubMed/NCBI
34	Dai H, Gao H, Zhao X, Dai L, Zhang X, Xiao N, Zhao R and Hemmingsen SM: Construction and characterization of a novel recombinant single-chain variable fragment antibody against white spot syndrome virus from shrimp. J Immunol Methods. 279:267–275. 2003. View Article : Google Scholar : PubMed/NCBI
35	Jäger M and Plückthun A: Domain interactions in antibody Fv and scFv fragments: Effects on unfolding kinetics and equilibria. FEBS Lett. 462:307–312. 1999. View Article : Google Scholar : PubMed/NCBI
36	Paoletti F, Malerba F, Konarev PV, Visintin M, Scardigli R, Fasulo L, Lamba D, Svergun DI and Cattaneo A: Direct intracellular selection and biochemical characterization of a recombinant anti-proNGF single chain antibody fragment. Arch Biochem Biophys. 522:26–36. 2012. View Article : Google Scholar : PubMed/NCBI
37	Adams GP and Schier R: Generating improved single-chain Fv molecules for tumor targeting. J Immunol Methods. 231:249–260. 1999. View Article : Google Scholar : PubMed/NCBI
38	Adams GP, Schier R, Marshall K, Wolf EJ, McCall AM, Marks JD and Weiner LM: Increased affinity leads to improved selective tumor delivery of single-chain Fv antibodies. Cancer Res. 58:485–490. 1998.PubMed/NCBI
39	Adams GP, Schier R, McCall AM, Simmons H, Horak E, Marks JD and Weiner LM: What are the determinants of antibody-based targeting? Proc Amer Assoc Cancer Res. 39:4361998.
40	Sela-Culang I, Kunik V and Ofran Y: The structural basis of antibody-antigen recognition. Front Immunol. 4:3022013. View Article : Google Scholar : PubMed/NCBI
41	Goldsby RA, Kindt TJ, Kuby J and Osborne BA: Immunology. 5th. W. H. Freeman and Company Publishers; New York: 2003
42	Burkovitz A, Leiderman O, Sela-Culang I, Byk G and Ofran Y: Computational identification of antigen-binding antibody fragments. J Immunol. 190:2327–2334. 2013. View Article : Google Scholar : PubMed/NCBI