Russian Journal of Biological Physics and Chemisrty Russian Journal of Biological Physics and Chemisrty АКТУАЛЬНЫЕ ВОПРОСЫ БИОЛОГИЧЕСКОЙ ФИЗИКИ И ХИМИИ 2499-9962 54347 МЕДИЦИНСКАЯ БИОФИЗИКА И БИОФИЗИЧЕСКАЯ ХИМИЯ MEDICAL BIOPHYSICS AND BIOPHYSICAL CHEMISTRY МЕДИЦИНСКАЯ БИОФИЗИКА И БИОФИЗИЧЕСКАЯ ХИМИЯ STRUCTURAL AND FUNCTIONAL PECULIARITIES OF a-CRYSTALLIN СТРУКТУРОНО-ФУНКЦИОНАЛЬНЫЕ ОСОБЕННОСТИ α-КРИСТАЛЛИНА Селиванова О М Selivanova O M seliv@vega.protres.ru Глякина А В Glyakina A V Суворина М Ю Suvorina M Yu Никулин А Д Nikulin A D Сурин А К Surin A K Галзитская О В Galzitskaya O V ogalzit@vega.protres.ru Институт белка РАН ru Institute of Protein Research, Russian Academy of Sciences ru Институт белка РАН ru Institute of Protein Research, Russian Academy of Sciences ru Институт белка РАН ru Institute of Protein Research, Russian Academy of Sciences ru Институт белка РАН ru Institute of Protein Research, Russian Academy of Sciences ru Институт белка РАН ru Institute of Protein Research, Russian Academy of Sciences ru Институт белка РАН ru Institute of Protein Research, Russian Academy of Sciences ru 25 09 2018 25 09 2018 3 3 673 679 20 09 2018 20 09 2018 https://rusjbpc.ru/en/nauka/article/54347/view

Α-Кристаллин является основным белком хрусталика глаза позвоночных. Под α-кристаллином понимается смесь гомологичных αА- и αВ- кристаллинов, которые формируют большие гетерогенные комплексы и отвечают за прозрачность хрусталика глаза. Установлено, что α-кристаллин принадлежит к семейству малых белков теплового шока (sHsp) и выполняет шапероно-подобную функцию, предотвращая денатурацию и агрегацию белков. 3D структура нативного α-кристаллина не установлена. В данной работе мы исследовали с помощью метода электронной микроскопии (ЭМ) структуру α-кристаллина из хрусталика глаза быка. Наши экспериментальные данные привели к выводу, что получить достоверную структуру α-кристаллинового комплекса невозможно. Анализ литературу по изучению α-кристаллина привел нас с заключению, что в нативных in vivo условиях структура и функционирование α-кристаллина может отличаться от его поведения в условиях in vitro . В связи с этим высказывается ряд предположений относительно функционирования α-кристаллина в виде небольших его агрегатов (например, димеров) и роли αА-кристаллина в качестве специализированного шаперона хрусталика глаза (шаперона для αВ-кристаллина).

A-Crystallin is the basic protein of the eye lens of vertebrates. It represents a mixture of homologous aA- and aB-crystallins which form large heterogeneous complexes and provide for eye lens transparency. It was established that a-crystallin belongs to the family of small heat shock proteins (sHsp) and performs a chaperone-like function preventing denaturation and aggregation of proteins. The 3D-structure of native a-crystallin has not yet been determined. In this study we have analyzed the structure of a-crystallin from bovine eye lens using the electron microscopy (EM) method. Our experimental data have allowed us to conclude that it is impossible to obtain a reliable structure of the a-crystallin complex. Based on the analysis of the publications devoted to studying a-crystallin we have come to the conclusion that under native in vivo conditions the structure and functioning of a-crystallin can differ from its behavior under in vitro conditions. Therefore several assumptions are made concerning the functioning of a-crystallin in the form of small aggregates, for example dimers, and the role of aA-crystallin as a specified chaperone of eye lens - the chaperone for aB-crystallin.

 α-кристаллин малые белки теплового шока электронная микроскопия a-crystallin small heat-shock proteins electron microscopy

Benedek G. Why the eye lens is transparent. Nature, 1983, vol. 302, pp. 383-384. Benedek G. Why the eye lens is transparent. Nature, 1983, vol. 302, pp. 383-384. Delaye M., Tardieu A. Short-range order of crystallin proteins accounts for eye lens transparency. Nature, 1983, vol. 302, pp. 415-417. Delaye M., Tardieu A. Short-range order of crystallin proteins accounts for eye lens transparency. Nature, 1983, vol. 302, pp. 415-417. Philipson B. Distribution of protein within lenses with x-ray cataract. Invest. Ophthalmol., 1969, vol. 8, pp. 271-280. Philipson B. Distribution of protein within lenses with x-ray cataract. Invest. Ophthalmol., 1969, vol. 8, pp. 271-280. Pierscionek B.K., Augusteyn R.C. The refractive index and protein distribution in the blue eye trevally lens. J. Am. Optom. Assoc., 1995, vol. 66, pp. 739-743. Pierscionek B.K., Augusteyn R.C. The refractive index and protein distribution in the blue eye trevally lens. J. Am. Optom. Assoc., 1995, vol. 66, pp. 739-743. Pierscionek B.K., Chan D.Y. Refractive index gradient of human lenses. Optom. Vis. Sci. Off. Publ. Am. Acad. Optom., 1989, vol. 66, pp. 822-829. Pierscionek B.K., Chan D.Y. Refractive index gradient of human lenses. Optom. Vis. Sci. Off. Publ. Am. Acad. Optom., 1989, vol. 66, pp. 822-829. Vérétout F., Delaye M., Tardieu A. Molecular basis of eye lens transparency. Osmotic pressure and X-ray analysis of alpha-crystallin solutions. J. Mol. Biol., 1989, vol. 205, pp. 713-728. Vérétout F., Delaye M., Tardieu A. Molecular basis of eye lens transparency. Osmotic pressure and X-ray analysis of alpha-crystallin solutions. J. Mol. Biol., 1989, vol. 205, pp. 713-728. Fagerholm P.P., Philipson B.T., Lindström B. Normal human lens - the distribution of protein. Exp. Eye Res., 1981, vol. 33, pp. 615-620. Fagerholm P.P., Philipson B.T., Lindström B. Normal human lens - the distribution of protein. Exp. Eye Res., 1981, vol. 33, pp. 615-620. Bloemendal H., de Jong W., Jaenicke R., Lubsen N.H., Slingsby C., Tardieu A. Ageing and vision: structure, stability and function of lens crystallins. Prog. Biophys. Mol. Biol., 2004, vol. 86, pp. 407-485. Bloemendal H., de Jong W., Jaenicke R., Lubsen N.H., Slingsby C., Tardieu A. Ageing and vision: structure, stability and function of lens crystallins. Prog. Biophys. Mol. Biol., 2004, vol. 86, pp. 407-485. de Jong W.W., Caspers G.J., Leunissen J.A. Genealogy of the alpha-crystallin-small heat-shock protein superfamily. Int. J. Biol. Macromol., 1998, vol. 22, pp. 151-162. de Jong W.W., Caspers G.J., Leunissen J.A. Genealogy of the alpha-crystallin-small heat-shock protein superfamily. Int. J. Biol. Macromol., 1998, vol. 22, pp. 151-162. Horwitz J. Alpha-crystallin. Exp. Eye Res., 2003, vol. 76, pp. 145-153. Horwitz J. Alpha-crystallin. Exp. Eye Res., 2003, vol. 76, pp. 145-153. Shiliaev N.G., Selivanova O.M., Galzitskaya O.V. Search for conserved amino acid residues of the α-crystallin proteins of vertebrates. J. Bioinform. Comput. Biol., 2016, vol. 14, p. 1641004. Shiliaev N.G., Selivanova O.M., Galzitskaya O.V. Search for conserved amino acid residues of the α-crystallin proteins of vertebrates. J. Bioinform. Comput. Biol., 2016, vol. 14, p. 1641004. Bloemendal H. The vertebrate eye lens. Science, 1977, vol. 197, pp. 127-138. Bloemendal H. The vertebrate eye lens. Science, 1977, vol. 197, pp. 127-138. Quax-Jeuken Y., Quax W., van Rens G., Khan P.M., Bloemendal H. Complete structure of the alpha B-crystallin gene: conservation of the exon-intron distribution in the two nonlinked alpha-crystallin genes. Proc. Natl. Acad. Sci. U.S.A., 1985, vol. 82, pp. 5819-5823. Quax-Jeuken Y., Quax W., van Rens G., Khan P.M., Bloemendal H. Complete structure of the alpha B-crystallin gene: conservation of the exon-intron distribution in the two nonlinked alpha-crystallin genes. Proc. Natl. Acad. Sci. U.S.A., 1985, vol. 82, pp. 5819-5823. Ngo J.T., Klisak I., Dubin R.A., Piatigorsky J., Mohandas T., Sparkes R.S., Bateman J.B. Assignment of the alpha B-crystallin gene to human chromosome 11. Genomics, 1989, vol. 5, pp. 665-669. Ngo J.T., Klisak I., Dubin R.A., Piatigorsky J., Mohandas T., Sparkes R.S., Bateman J.B. Assignment of the alpha B-crystallin gene to human chromosome 11. Genomics, 1989, vol. 5, pp. 665-669. Srinivasan A.N., Nagineni C.N., Bhat S.P. Alpha A-crystallin is expressed in non-ocular tissues. J. Biol. Chem., 1992, vol. 267, pp. 23337-23341. Srinivasan A.N., Nagineni C.N., Bhat S.P. Alpha A-crystallin is expressed in non-ocular tissues. J. Biol. Chem., 1992, vol. 267, pp. 23337-23341. Renkawek K., de Jong W.W., Merck K.B., Frenken C.W., van Workum F.P., Bosman G.J. Alpha B-crystallin is present in reactive glia in Creutzfeldt-Jakob disease. Acta Neuropathol. (Berl.), 1992, vol. 83, pp. 324-327. Renkawek K., de Jong W.W., Merck K.B., Frenken C.W., van Workum F.P., Bosman G.J. Alpha B-crystallin is present in reactive glia in Creutzfeldt-Jakob disease. Acta Neuropathol. (Berl.), 1992, vol. 83, pp. 324-327. Renkawek K., Voorter C.E., Bosman G.J., van Workum F.P., de Jong W.W. Expression of alpha B-crystallin in Alzheimer’s disease. Acta Neuropathol. (Berl.), 1994, vol. 87, pp. 155-160. Renkawek K., Voorter C.E., Bosman G.J., van Workum F.P., de Jong W.W. Expression of alpha B-crystallin in Alzheimer’s disease. Acta Neuropathol. (Berl.), 1994, vol. 87, pp. 155-160. Horwitz J. Alpha-crystallin can function as a molecular chaperone. Proc. Natl. Acad. Sci. U.S.A., 1992, vol. 89, pp. 10449-10453. Horwitz J. Alpha-crystallin can function as a molecular chaperone. Proc. Natl. Acad. Sci. U.S.A., 1992, vol. 89, pp. 10449-10453. Horwitz J., Huang Q.L., Ding L., Bova M.P. Lens alpha-crystallin: chaperone-like properties. Methods Enzymol., 1998, vol. 290, pp. 365-383. Horwitz J., Huang Q.L., Ding L., Bova M.P. Lens alpha-crystallin: chaperone-like properties. Methods Enzymol., 1998, vol. 290, pp. 365-383. Siezen R.J., Berger H. The quaternary structure of bovine alpha-crystallin. Size and shape studies by sedimentation, small-angle X-ray scattering and quasi-elastic light scattering. Eur. J. Biochem., 1978, vol. 91, pp. 397-405. Siezen R.J., Berger H. The quaternary structure of bovine alpha-crystallin. Size and shape studies by sedimentation, small-angle X-ray scattering and quasi-elastic light scattering. Eur. J. Biochem., 1978, vol. 91, pp. 397-405. de Jong W.W., Caspers G.J., Leunissen J.A. Genealogy of the alpha-crystallin-small heat-shock protein superfamily. Int. J. Biol. Macromol., 1998, vol. 22, pp. 151-162. de Jong W.W., Caspers G.J., Leunissen J.A. Genealogy of the alpha-crystallin-small heat-shock protein superfamily. Int. J. Biol. Macromol., 1998, vol. 22, pp. 151-162. Haslbeck M., Peschek J., Buchner J., Weinkauf S. Structure and function of α-crystallins: Traversing from in vitro to in vivo. Biochim. Biophys. Acta, 2016, vol. 1860, pp. 149-166. Haslbeck M., Peschek J., Buchner J., Weinkauf S. Structure and function of α-crystallins: Traversing from in vitro to in vivo. Biochim. Biophys. Acta, 2016, vol. 1860, pp. 149-166. Peschek J., Braun N., Franzmann T.M., Georgalis Y., Haslbeck M., Weinkauf S., Buchner J. The eye lens chaperone alpha-crystallin forms defined globular assemblies. Proc. Natl. Acad. Sci. U.S.A., 2009, vol. 106, pp. 13272-13277. Peschek J., Braun N., Franzmann T.M., Georgalis Y., Haslbeck M., Weinkauf S., Buchner J. The eye lens chaperone alpha-crystallin forms defined globular assemblies. Proc. Natl. Acad. Sci. U.S.A., 2009, vol. 106, pp. 13272-13277. Kim K.K., Kim R., Kim S.H. Crystal structure of a small heat-shock protein. Nature, 1998, vol. 394, pp. 595-599. Kim K.K., Kim R., Kim S.H. Crystal structure of a small heat-shock protein. Nature, 1998, vol. 394, pp. 595-599. Haley D.A., Horwitz J., Stewart P.L. The small heat-shock protein, alphaB-crystallin, has a variable quaternary structure. J. Mol. Biol., 1998, vol. 277, pp. 27-35. Haley D.A., Horwitz J., Stewart P.L. The small heat-shock protein, alphaB-crystallin, has a variable quaternary structure. J. Mol. Biol., 1998, vol. 277, pp. 27-35. Haley D.A., Bova M.P., Huang Q.L., Mchaourab H.S., Stewart P.L. Small heat-shock protein structures reveal a continuum from symmetric to variable assemblies. J. Mol. Biol., 2000, vol. 298, pp. 261-272. Haley D.A., Bova M.P., Huang Q.L., Mchaourab H.S., Stewart P.L. Small heat-shock protein structures reveal a continuum from symmetric to variable assemblies. J. Mol. Biol., 2000, vol. 298, pp. 261-272. Jehle S., Vollmar B.S., Bardiaux B., Dove K.K., Rajagopal P., Gonen T., Oschkinat H., Klevit R.E. N-terminal domain of alphaB-crystallin provides a conformational switch for multimerization and structural heterogeneity. Proc. Natl. Acad. Sci. U.S.A., 2011, vol. 108, pp. 6409-6414. Jehle S., Vollmar B.S., Bardiaux B., Dove K.K., Rajagopal P., Gonen T., Oschkinat H., Klevit R.E. N-terminal domain of alphaB-crystallin provides a conformational switch for multimerization and structural heterogeneity. Proc. Natl. Acad. Sci. U.S.A., 2011, vol. 108, pp. 6409-6414. Braun N., Zacharias M., Peschek J., Kastenmüller A., Zou J., Hanzlik M., Haslbeck M., Rappsilber J., Buchner J., Weinkauf S. Multiple molecular architectures of the eye lens chaperone αB-crystallin elucidated by a triple hybrid approach. Proc. Natl. Acad. Sci. U.S., 2011, vol. 108, pp. 20491-20496. Braun N., Zacharias M., Peschek J., Kastenmüller A., Zou J., Hanzlik M., Haslbeck M., Rappsilber J., Buchner J., Weinkauf S. Multiple molecular architectures of the eye lens chaperone αB-crystallin elucidated by a triple hybrid approach. Proc. Natl. Acad. Sci. U.S., 2011, vol. 108, pp. 20491-20496. Peschek J., Braun N., Rohrberg J., Back K.C., Kriehuber T., Kastenmuller A., Weinkauf S., Buchner J. Regulated structural transitions unleash the chaperone activity of αB-crystallin. Proc. Natl. Acad. Sci., 2013, vol. 110, E3780-E3789. Peschek J., Braun N., Rohrberg J., Back K.C., Kriehuber T., Kastenmuller A., Weinkauf S., Buchner J. Regulated structural transitions unleash the chaperone activity of αB-crystallin. Proc. Natl. Acad. Sci., 2013, vol. 110, E3780-E3789. Kampinga H.H., Garrido C. HSPBs: small proteins with big implications in human disease. Int. J. Biochem. Cell Biol., 2012, vol. 44, pp. 1706-1710. Kampinga H.H., Garrido C. HSPBs: small proteins with big implications in human disease. Int. J. Biochem. Cell Biol., 2012, vol. 44, pp. 1706-1710. Chiou S.H., Azari P., Himmel M.E., Squire P.G. Isolation and physical characterization of bovine lens crystallins. Int. J. Pept. Protein Res., 1979, vol. 13, pp. 409-417. Chiou S.H., Azari P., Himmel M.E., Squire P.G. Isolation and physical characterization of bovine lens crystallins. Int. J. Pept. Protein Res., 1979, vol. 13, pp. 409-417. Spector A., Li L.K., Augusteyn R.C., Schneider A., Freund T. α-Crystallin. The isolation and characterization of distinct macromolecular fractions. Biochem. J., 1971, vol. 124, pp. 337-343. Spector A., Li L.K., Augusteyn R.C., Schneider A., Freund T. α-Crystallin. The isolation and characterization of distinct macromolecular fractions. Biochem. J., 1971, vol. 124, pp. 337-343. Siezen R.J., Bindels J.G., Hoenders H.J. The quaternary structure of bovine alpha-crystallin. Size and charge microheterogeneity: more than 1000 different hybrids? Eur. J. Biochem., 1978, vol. 91, pp. 387-396. Siezen R.J., Bindels J.G., Hoenders H.J. The quaternary structure of bovine alpha-crystallin. Size and charge microheterogeneity: more than 1000 different hybrids? Eur. J. Biochem., 1978, vol. 91, pp. 387-396. Bloemendal H., Berns T., Zweers A., Hoenders H., Benedetti E.L. The state of aggregation of α-crystallin detected after large-scale preparation by zonal centrifugation. Eur. J. Biochem., 1972, vol. 24, pp. 401-406. Bloemendal H., Berns T., Zweers A., Hoenders H., Benedetti E.L. The state of aggregation of α-crystallin detected after large-scale preparation by zonal centrifugation. Eur. J. Biochem., 1972, vol. 24, pp. 401-406. Peschek J., Braun N., Rohrberg J., Back K.C., Kriehuber T., Kastenmüller A., Weinkauf S., Buchner J. Regulated structural transitions unleash the chaperone activity of αB-crystallin. Proc. Natl. Acad. Sci. U.S.A., 2013, vol. 110, pp. E3780-3789. Peschek J., Braun N., Rohrberg J., Back K.C., Kriehuber T., Kastenmüller A., Weinkauf S., Buchner J. Regulated structural transitions unleash the chaperone activity of αB-crystallin. Proc. Natl. Acad. Sci. U.S.A., 2013, vol. 110, pp. E3780-3789. Gu L., Abulimiti A., Li W., Chang Z. Monodisperse Hsp16.3 Nonamer Exhibits Dynamic Dissociation and Reassociation, with the Nonamer Dissociation Prerequisite for Chaperone-like Activity. J. Mol. Biol., vol. 319, pp. 517-526. Gu L., Abulimiti A., Li W., Chang Z. Monodisperse Hsp16.3 Nonamer Exhibits Dynamic Dissociation and Reassociation, with the Nonamer Dissociation Prerequisite for Chaperone-like Activity. J. Mol. Biol., vol. 319, pp. 517-526. Brady J.P., Garland D., Duglas-Tabor Y., Robison W.G., Jr. Groome A., Wawrousek E.F. Targeted disruption of the mouse alpha A-crystallin gene induces cataract and cytoplasmic inclusion bodies containing the small heat shock protein alpha B-crystallin. Proc. Natl. Acad. Sci. U.S.A., 1997, vol. 94, pp. 884-889. Brady J.P., Garland D., Duglas-Tabor Y., Robison W.G., Jr. Groome A., Wawrousek E.F. Targeted disruption of the mouse alpha A-crystallin gene induces cataract and cytoplasmic inclusion bodies containing the small heat shock protein alpha B-crystallin. Proc. Natl. Acad. Sci. U.S.A., 1997, vol. 94, pp. 884-889. Ma Z., Hanson S.R.A., Lampi K.J., David L.L., Smith D.L., Smith J.B. Age-Related Changes in Human Lens Crystallins Identified by HPLC and Mass Spectrometry. Exp. Eye Res., 1998, vol. 67, pp. 21-30. Ma Z., Hanson S.R.A., Lampi K.J., David L.L., Smith D.L., Smith J.B. Age-Related Changes in Human Lens Crystallins Identified by HPLC and Mass Spectrometry. Exp. Eye Res., 1998, vol. 67, pp. 21-30.