Shriners Research Center, Portland OR

Bächinger Lab
Research interest: 
Our major interest is in the biosynthesis and folding of procollagens. Extensive posttranslational modifications involving different resident ER enzymes are required for proper collagen assembly and folding. Among those enzymes are collagen prolyl hydroxylases, namely well known prolyl-4-hydroxylase and recently characterized in our lab prolyl-3-hydroxylase. Both of these enzymes hydroxylate proline residues in collagens, but do this with different frequency and in different positions. In all types of collagen 4-hydroxylation of proline residues in the Yaa position is crucial for the formation of triple helix. Almost all prolines in Yaa position of the Gly-Xaa-Yaa repeated sequence are hydroxylated to 4(R)-hydroxyproline by prolyl-4-hydroxylase. The role of 3(S)-hydroxyproline in the Xaa position of Gly-Xaa-Yaa repeated sequence of collagens accomplished by prolyl-3-hydroxylase is not well understood. The extent of 3-hydroxylation varies with different types of collagen and occurs in largest amount in collagen types IV and V. Little is known about the structure of prolyl-3-hydroxylase. Analysis of the primary sequence of prolyl-3-hydroxylases identifies two major domains: the 2-oxoglutarate- and Fe(II)-dependent dioxygenase carboxy-terminal domain and conserved CXXXC repeating motif in the amino-terminal domain. It was also demonstrated that prolyl-3-hydroxylase 1 (P3H1) exists as a stable complex of three proteins P3H1, CRTAP and CycB in 1:1:1 ratio. Interestingly, the amino acid sequence of CRTAP is significantly homologous to that of N-terminal domain of P3H1 and both proteins have the same number of CXXXC repeats. These cysteine-rich motifs can potentially exhibit a disulfide isomerase-like activity. The P3H1/CRTAP/CypB complex has been shown to be very important for the biosynthesis of procollagens. Human mutations in the genes for P3H1 and CRTAP lead to severe Osteogenesis Imperfecta (OI). Elucidation of the three-dimensional structure of P3H1/CRTAP/CypB complex is a major interest of my current research.

BIOGRAPHICAL SKETCH
Name: Elena N. Pokidysheva
Role on Project: Postdoc

Institution & Location                                   Degree       Year(s)              Field of Study
Moscow Institute of Physics and Technology    Diploma      1999                  Mathematics & Physics
University of California Santa Barbara               N/A      2000-2001             Biophysics
University of Basel                                          Ph.D.         2004                 Biophysical Chemistry.

Research and/or Professional Experience:
2006-Present Postdoc with Dr.Hans Peter Bächinger, Shriners Hospitals for Children, OHSU, Portland,Oregon
2004-2006 Postdoctoral Fellow with Professor Michael Rossmann, Purdue University, West Lafayette, Indiana
2002-2004 Ph.D. student with Professor Jurgen Engel, Biozentrum, University of Basel, Basel, Switzerland
2000-2001 Ph.D. student with Professor Cyrus Safinya, University of California Santa Barbara, Goleta, California
1995-2000 Diploma student with Professor Sevastianov V.I., Center for Blood Compatible Biomaterials, Research Inst. of Transplantology and Artificial Organs, Moscow, Russian Federation

Peer-Reviewed Publications (published, in press and submitted):
Pokidysheva E, Zhang Y, Battisti AJ, Bator-Kelly CM, Chipman PR, Xiao C, Gregorio GG, Hendrickson WA, Kuhn RJ, Rossmann MG., Cryo-EM reconstruction of dengue virus in complex with the carbohydrate recognition domain of DC-SIGN. Cell. 2006 Feb 10;124(3):485-93
AG, Moroder L, Engel J, Holstein TW., The glycoprotein NOWA and minicollagens are part of a disulfide-linked polymer that forms the cnidarian nematocyst wall. J Biol Chem. 2004 Dec 10;279(50):52016-23. Epub 2004 Oct 4.
Pokidysheva E, Milbradt AG, Meier S, Renner C, Haussinger D, BachingerHP, Moroder L, Grzesiek S, Holstein TW, Ozbek S, Engel J., The structure of the Cys-rich terminal domain of Hydra minicollagen, which isinvolved in disulfide networks of the nematocyst wall. J Biol Chem. 2004 Jul 16;279(29):30395-401.
Meier S, Haussinger D, Pokidysheva E, Bachinger HP, Grzesiek S., Determination of a high-precision NMR structure of the minicollagen cysteine rich domain from Hydra and characterization of its disulfide bond formation. FEBS Lett. 2004 Jul 2;569(1-3):112-6.
O. Pelltier, E. Pokidysheva, L.S. Hirst, N. Bouxsein, Y. Li, and C. R. Safinya, Structure of actin cross-linked with alpha-actinin: a network of bundles PHYS. REV. LETT. 91, 148102 Sept 2003
E.N. Pokidysheva; I.A. Maklakova; Z.M. Belomestnaya; N.V. Perova; S.N.Bagrov; V.I. Sevastianov., Comparative analysis of human serum albumin adsorption and complement activation for hydrophobic and hydrophilic intraocular lenses. Artif Organs. 2001 Jun;25(6):453-8.
S. Petrash, T. Cregger, B. Zhao, E. Pokidysheva, M. D. Foster, W. J. Brittain, V. Sevastianov and C. F. Majkrzak, Changes in Protein Adsorption on Self-Assembled Monolayers with Monolayer Order: Comparison of Human Serum Albumin and Human Gamma Globulin. Langmuir 2001, 17, 7645-7651.
Titushkin, IA; Vasin, SL; Rozanova, IB; Pokidysheva, EN; Alekhin, AP; Sevastianov,VI., Carbon coated polyethylene: Effect of surface energetics and topography on human platelet adhesion. ASAIO JOURNAL, JAN-FEB, 2001, V47(N1):11-17.
Pokidysheva, EN; Nemets, EA; Tremsina, YS; Sevast'yanov, VI., Competitive adsorption of human fibrinogen on the surface of amorphous quartz. BIOFIZIKA, SEP-OCT, 2000, V45(N5):809-815.
V.I. Sevastianov; E.N. Pokidysheva; I.A. Maklakova; S.N. Bagrov., Use of the fluorescence method in the study of protein adsorption onto the surface of intraocular lenses. Medical Engineering, 1998 Sep-Oct,(5):3-6.