Cataract-causing mutation S228P promotes βB1-crystallin aggregation and degradation by separating two interacting loops in C-terminal domain

Liang-Bo Qi, Li-Dan Hu, Huihui Liu, Hai-Yun Li, Xiao-Yao Leng, Yong-Bin Yan

PDF(2785 KB)
PDF(2785 KB)
Protein Cell ›› 2016, Vol. 7 ›› Issue (7) : 501-515. DOI: 10.1007/s13238-016-0284-3
RESEARCH ARTICLE

Cataract-causing mutation S228P promotes βB1-crystallin aggregation and degradation by separating two interacting loops in C-terminal domain

Author information +
History +

Abstract

β/γ-Crystallins are predominant structural proteins in the cytoplasm of lens fiber cells and share a similar fold composing of four Greek-key motifs divided into two domains. Numerous cataract-causing mutations have been identified in various β/γ-crystallins, but the mechanisms underlying cataract caused by most mutations remains uncharacterized. The S228P mutation in βB1-crystallin has been linked to autosomal dominant congenital nuclear cataract. Here we found that the S228P mutant was prone to aggregate and degrade in both of the human and E. coli cells. The intracellular S228P aggregates could be redissolved by lanosterol. The S228P mutation modified the refolding pathway of βB1-crystallin by affecting the formation of the dimeric intermediate but not the monomeric intermediate. Compared with native βB1-crystallin, the refolded S228P protein had less packed structures, unquenched Trp fluorophores and increased hydrophobic exposure. The refolded S228P protein was prone to aggregate at the physiological temperature and decreased the protective effect of βB1-crystallin on βA3-crystallin. Molecular dynamic simulation studies indicated that the mutation decreased the subunit binding energy and modified the distribution of surface electrostatic potentials. More importantly, the mutation separated two interacting loops in the C-terminal domain, which shielded the hydrophobic core from solvent in native βB1-crystallin. These two interacting loops are highly conserved in both of the N- and C-terminal domains of all β/γ-crystallins. We propose that these two interacting loops play an important role in the folding and structural stability of β/γ-crystallin domains by protecting the hydrophobic core from solvent access.

Keywords

β/γ-crystallin / cataract-causing mutation / hydrophobic core / protein aggregation / protein folding

Cite this article

Download citation ▾
Liang-Bo Qi, Li-Dan Hu, Huihui Liu, Hai-Yun Li, Xiao-Yao Leng, Yong-Bin Yan. Cataract-causing mutation S228P promotes βB1-crystallin aggregation and degradation by separating two interacting loops in C-terminal domain. Protein Cell, 2016, 7(7): 501‒515 https://doi.org/10.1007/s13238-016-0284-3

References

[1]
Andley UP (2007) Crystallins in the eye: function and pathology. Prog Retin Eye Res. 26:78–98
CrossRef Google scholar
[2]
Baldwin RL (2007) Energetics of protein folding. J Mol Biol 371:283–301
CrossRef Google scholar
[3]
Bassnett S (2009) On the mechanism of organelle degradation in the vertebrate lens. Exp Eye Res 88:133–139
CrossRef Google scholar
[4]
Bateman OA, Lubsen NH, Slingsby C (2001) Association behaviour of human βB1-crystallin and its truncated forms. Exp Eye Res 73:321–331
CrossRef Google scholar
[5]
Bateman OA, Sarra R, van Genesen ST, Kapp G, Lubsen NH, Slingsby C (2003) The stability of human acidic β-crystallin oligomers and hetero-oligomers. Exp Eye Res 77:409–422
CrossRef Google scholar
[6]
Bax B, Lapatto R, Nalini V, Driessen H, Lindley PF, Mahadevan D, Blundell TL, Slingsby C (1990) X-ray analysis of βB2-crystallin and evolution of oligomeric lens proteins. Nature 347:776–780
CrossRef Google scholar
[7]
Benedek GB (1971) Theory of transparency of the eye. Appl Opt 10:459–473
CrossRef Google scholar
[8]
Benedek GB (1997) Cataract as a protein condensation disease: the Proctor Lecture. Invest Ophthalmol Vis Sci 38:1911–1921
[9]
Berbers GA, Hoekman WA, Bloemendal H, de Jong WW, Kleinschmidt T, Braunitzer G (1983) Proline- and alanine-rich N-terminal extension of the basic bovine β-crystallin B1 chains. FEBS Lett 161:225–229
CrossRef Google scholar
[10]
Bloemendal H, de Jong W, Jaenicke R, Lubsen NH, Slingsby C, Tardieu A (2004) Ageing and vision: structure, stability and function of lens crystallins. Prog Biophys Mol Biol 86:407–485
CrossRef Google scholar
[11]
Chen J, Flaugh SL, Callis PR, King J (2006) Mechanism of the highly efficient quenching of tryptophan fluorescence in human γDcrystallin. Biochemistry 45:11552–11563
CrossRef Google scholar
[12]
Chen J, Callis PR, King J (2009) Mechanism of the very efficient quenching of tryptophan fluorescence in human γD- and γScrystallins: the γ-crystallin fold may have evolved to protect tryptophan residues from ultraviolet photodamage. Biochemistry 48:3708–3716
CrossRef Google scholar
[13]
Chen Z, Chen X-J, Xia M, He H-W, Wang S, Liu H, Gong H, Yan Y-B (2012) Chaperone-like effect of the linker on the isolated C-terminal domain of rabbit muscle creatine kinase. Biophys J 103:558–566
CrossRef Google scholar
[14]
Chennamsetty N, Voynov V, Kayser V, Helk B, Trout BL (2009) Design of therapeutic proteins with enhanced stability. Proc Natl Acad Sci USA 106:11937–11942
CrossRef Google scholar
[15]
Collins TJ (2007) ImageJ for microscopy. Biotechniques 43:25–30
CrossRef Google scholar
[16]
David LL, Lampi KJ, Lund AL, Smith JB (1996) The sequence of human bB1-crystallin cDNA allows mass spectrometric detection of βB1 protein missing portions of its N-terminal extension. J Biol Chem 271:4273–4279
CrossRef Google scholar
[17]
Dolinska MB, Sergeev YV, Chan MP, Palmer I, Wingfield PT (2009) N-terminal extension of βB1-crystallin: identification of a critical region that modulates protein interaction with βA3-crystallin. Biochemistry 48(40):9684–9695
[18]
Flaugh SL, Kosinski-Collins MS, King J (2005) Contributions of hydrophobic domain interface interactions to the folding and stability of human γD-crystallin. Protein Sci 14:569–581
CrossRef Google scholar
[19]
He G-J, Zhang A, Liu W-F, Cheng Y, Yan Y-B (2009) Conformational stability and multistate unfolding of poly(A)-specific ribonuclease. FEBS J 276:2849–2860
CrossRef Google scholar
[20]
Horwitz J (1992) α-Crystallin can function as a molecular chaperone. Proc Natl Acad Sci USA 89:10449–10453
CrossRef Google scholar
[21]
Kosinski-Collins MS, Flaugh SL, King J (2004) Probing folding and fluorescence quenching in human γD crystallin Greek key domains using triple tryptophan mutant proteins. Protein Sci 13:2223–2235
CrossRef Google scholar
[22]
Kroone RC, Elliott GS, Ferszt A, Slingsby C, Lubsen NH, Schoenmakers JG (1994) The role of the sequence extensions in β-crystallin assembly. Protein Eng 7:1395–1399
CrossRef Google scholar
[23]
Leng XY, Wang S, Cao NQ, Qi LB, Yan YB (2014) The N-terminal extension of βB1-crystallin chaperones β-crystallin folding and cooperates with αA-crystallin. Biochemistry 53:2464–2473
CrossRef Google scholar
[24]
Lin H, Ouyang H, Zhu J, Huang S, Liu Z, Chen S, Cao G, Li G, Signer RA, Xu Y, Chung C, Zhang Y, Lin D, Patel S, Wu F, Cai H, Hou J, Wen C, Jafari M, Liu X, Luo L, Zhu J, Qiu A, Hou R, Chen B, Chen J, Granet D, Heichel C, Shang F, Li X, Krawczyk M, Skowronska-Krawczyk D, Wang Y, Shi W, Chen D, Zhong Z, Zhong S, Zhang L, Chen S, Morrison SJ, Maas RL, Zhang K, Liu Y (2016) Lens regeneration using endogenous stem cells with gain of visual function. Nature 531:323–328
CrossRef Google scholar
[25]
Liu BF, Liang JJ (2005) Interaction and biophysical properties of human lens Q155* βB2-crystallin mutant. Mol Vis 11:321–327
[26]
Montfort RLMV, Bateman OA, Lubsen NH, Slingsby C (2003) Crystal structure of truncated human βB1-crystallin. Protein Sci. 12:2606–2612
[27]
Moreau KL, King J (2009) Hydrophobic core mutations associated with cataract development in mice destabilize human γDcrystallin. J Biol Chem 284:33285–33295
CrossRef Google scholar
[28]
Moreau KL, King JA (2012) Protein misfolding and aggregation in cataract disease and prospects for prevention. Trends Mol Med 18:273–282
CrossRef Google scholar
[29]
Morozov V, Wawrousek EF (2005) Arrested apoptosis in lens fiber cells: a possible role of α-crystallin. FEBS J 272:40
[30]
Phillips JC, Braun R, Wang W, Gumbart J, Tajkhorshid E, Villa E, Chipot C, Skeel RD, Kale L, Schulten K (2005) Scalable molecular dynamics with NAMD. J Comput Chem 26:1781–1802
CrossRef Google scholar
[31]
Prabhu NV, Sharp KA (2005) Heat capacity in proteins. Annu Rev Phys Chem 56:521–548
CrossRef Google scholar
[32]
Serebryany E, King JA (2014) The βγ-crystallins: native state stability and pathways to aggregation. Prog Biophys Mol Biol 115:32–41
CrossRef Google scholar
[33]
Sergeev YV, David LL, Chen HC, Hope JN, Hejtmancik JF (1998) Local microdomain structure in the terminal extensions of βA3-and βB2-crystallins. Mol Vis 4:9
[34]
Shiels A, Hejtmancik JF (2015) Molecular Genetics of Cataract. Prog Mol Biol Transl Sci. 134:203–218
CrossRef Google scholar
[35]
Slingsby C, Bateman OA (1990) Quaternary interactions in eye lens β-crystallins: basic and acidic subunits of β-crystallins favor heterologous association. Biochemistry 29:6592–6599
CrossRef Google scholar
[36]
Turoverov KK, Haitlina SY, Pinaev GP (1976) Ultra-violet fluorescence of actin. Determination of native actin content in actin preparations. FEBS Lett 62:4–6
CrossRef Google scholar
[37]
Vendra VPR, Agarwal G, Chandani S, Talla V, Srinivasan N, Balasubramanian D (2013) Structural integrity of the Greek key motif in βγ-crystallins is vital for central eye lens transparency. PLoS One 8:e70336
[38]
Wang J, Ma X, Gu F, Liu NP, Hao XL, Wang KJ, Wang NL, Zhu SQ (2007) A missense mutation S228P in the CRYBB1 gene causes autosomal dominant congenital cataract. Chin Med J (Engl) 120:820–824
[39]
Wang S, Leng XY, Yan YB (2011a) The benefits of being β-crystallin heteromers: βB1-crystallin protects βA3-crystallin against aggregation during co-refolding. Biochemistry 50:10451–10461
[40]
Wang KJ, Wang S, Cao NQ, Yan YB, Zhu SQ (2011b) A novel mutation in CRYBB1 associated with congenital cataract-microcornea syndrome: the p.Ser129Arg mutation destabilizes the βB1/βA3-crystallin heteromer but not the βB1-crystallin homomer. Hum Mutat 32:E2050–E2060
[41]
Wang S, Zhao WJ, Liu H, Gong H, Yan YB (2013) Increasing βB1-crystallin sensitivity to proteolysis caused by the congenital cataract-microcornea syndrome mutation S129R. Biochim Biophys Acta 1832:302–311
CrossRef Google scholar
[42]
Xi YB, Zhao WJ, Zuo XT, Tjondro JHC AB, Li S, Dai YB, WangS, YanYB (2014a) Cataract-causing mutation R233H affects the stabilities of βB1- and βA3/βB1-crystallins with different pH-dependence. Biochim Biophys Acta 1842:2216–2229
[43]
Xi YB, Zhang K, Dai AB, Ji SR, Yao K, Yan YB (2014b) Cataractlinked mutation R188H promotes βB2-crystallin aggregation and fibrillization during acid denaturation. Biochem Biophys Res Commun. 447:244–249
[44]
Xi YB, Chen XJ, Zhao WJ, Yan YB (2015) Congenital Cataractcausing mutation G129C in γC-crystallin promotes the accumulation of two distinct unfolding intermediates that form highly toxic aggregates. J Mol Biol 427:2765–2781
CrossRef Google scholar
[45]
Xu J, Wong C, Tan X, Jing H, Zhou G, Song W (2010) Decreasing the homodimer interaction: a common mechanism shared by the ΔG91 mutation and deamidation in βA3-crystallin. Mol Vis 16:438–444
[46]
Xu J, Wang S, Zhao WJ, Xi YB, Yan YB, Yao K (2012) The congenital cataract-linked A2V mutation impairs tetramer formation and promotes aggregation of βB2-crystallin. PLoS One 7: e51200
[47]
Zhang K, Zhao WJ, Leng XY, Wang S, Yao K, Yan YB (2014) The importance of the last strand at the C-terminus in βB2-crystallin stability and assembly. Biochim Biophys Acta 1842:44–55
CrossRef Google scholar
[48]
Zhao L, Chen X-J, Zhu J, Xi Y-B, Yang X, Hu L-D, Ouyang H, Patel SH, Jin X, Lin D, Wu F, Flagg K, Cai H, Li G, Cao G, Lin Y, Chen D, Wen C, Chung C, Wang Y, Qiu A, Yeh E, Wang W, Hu X, Grob S, Abagyan R, Su Z, Tjondro HC, Zhao X-J, Luo H, Hou R, Jefferson J, Perry P, Gao W, Kozak I, Granet D, Li Y, Sun X, Wang J, Zhang L, Liu Y, Yan Y-B, Zhang K (2015) Lanosterol reverses protein aggregation in cataracts. Nature 523:607–611
CrossRef Google scholar

RIGHTS & PERMISSIONS

2016 The Author(s) 2016. This article is published with open access at Springerlink.com and journal.hep.com.cn
AI Summary AI Mindmap
PDF(2785 KB)

Accesses

Citations

Detail

Sections
Recommended

/