Dr. Andreas Holzenburg
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Dr. Andreas Holzenburg Microscopy and Imaging Center
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Research Interests
My research group is employing electron microscopy (EM)/crystallography as a tool for the structural elucidation of (sub)cellular components and biological macromolecules, particularly transmembrane and membrane-associated proteins The structure determination involves image processing routines and is carried out in conjunction with thorough biochemical characterisations. Furthermore, we are using EM not only in isolation but also jointly with other biophysical approaches including X-ray crystallography and spectroscopical methods. As EM is an extremely versatile tool, a number of collaborations on structure-oriented projects have already benefited from the use of EM techniques:
- Blood coagulation factors (G Kemball-Cook, Hammersmith Hospital, London, UK)
- Parasitic nematode feeding tube assembly (with HJ Atkinson, Leeds, UK)
- Structure-function relationships of photosystem II (with RC Ford, UMIST, UK)
- Structure and function of the internal ribosomal entry site of the hepatitis C virus genome (with DJ Rowlands, Leeds, UK)
- Lon protease (with CD Thomas, Leeds, UK)
- Photosystem I and II from higher plants (with RC Ford, UMIST, UK)
- GABA receptor (with HH Xue, Hong Kong)
- RhD protein (with J-P Cartron/O Bertrand, INSERM-U76, Paris, France)
- p130 (with E Bogner, Marburg, FR Germany)
Structure and Function of the Plastid Division Machinery
The Holzenburg lab is interested in the structure, assembly and turnover dynamics of the cytoskeletal tubulin-like FtsZ protein in plant chloroplats. FtsZ plays a pivotal role in the division of prokaryotic cells as well as chloroplasts. One of the functional characteristics of FtsZ is its auto-assembly into a ring-like macromolecular complex called the Z-ring. During cell division, the Z-ring undergoes continuous and rapid remodeling via subunit exchange and constricts at the leading edge of the septum with the simultaneous loss of subunits. Consistent with the endosymbiotic origin of chloroplasts, plants possess nuclear-encoded, plastid-targeted homologues of bacterial FtsZ. However, whereas most prokaryotes, including the cyanobacterial relatives of chloroplasts, have a single form of FtsZ, two structurally distinct FtsZ protein families, FtsZ1 and FtsZ2. Experimental evidence suggests that FtsZ1 and FtsZ2 function in a complex and that their roles are functionally distinct. They are both localized in the stromal compartment of the chloroplast and are always tightly colocalized in immuno-fluorescence labeling experiments. In addition, FtsZ1 seems unable to form long polymers in planta without FtsZ2.
Overall, the molecular mechanism of FtsZ filament assembly and its regulation, the structures of assembled protofilaments, and the structure of the in vivo FtsZ ring in chloroplasts remain poorly understood. Our overall goal is to expand the current model for FtsZ assembly and investigate and define the molecular structure and assembly dynamics of FtsZ rings in chloroplasts of Arabidopsis thaliana with the view to understand chloroplast size control.
Modulation of the size of storage plastids (amyloplasts) and the starch granule size by changing the levels of FtsZ expression is of considerable interest to the starch and biofuel industry, since increased starch granule size improves the wet-milling efficiency and thus the starch yield in staple crops.
Dr. Holzenburg - Research Interests
Biography
My research background is in Microbiology, Biochemistry and Structural Biology. Throughout my postgraduate and postdoctoral research I have used 2D/3D crystallisation, electron microscopy [EM] and digital/analogue image analysis/processing in connection with biochemical as well as immunological techniques to study structure-function relationships of soluble and membrane proteins of bacterial, plant, animal, viral and human origin. During my stay in the USA I was concerned with the biochemistry and EM of Clostridium species and affiliated thermostable cellulases.
From 1-Jan-1991 I took up the appointment as lecturer in Structural Molecular Biology at the University of Leeds. This appointment was aimed at building up a vibrant EM-based research programme. I have trained and advised numerous individuals ranging from students to senior academics (intra- and extramural, national and international) in the relevant EM techniques, introduced analogue and digital image analysis/processing/display systems (CRISP, MRC, SPECTRA, SPIDER, SYNU), and established a cryo-electron microscopy facility. Since my Leeds appointment, I have mainly worked on membrane proteins (see also list of recent publications) but was also able to maintain a steady interest in more cellular/subcellular EM work resulting in the production of the RMS Handbook “Electron Microscopy in Microbiology” (bios Publishers Ltd., Oxford).
(1) March 1980- Dec.1983 Undergraduate research assistant (part-time) at the Max-Planck Institute for Exp. Medicine (with Prof. Dr. G. v. Ehrenstein, Drs. R.C. Cassada, K. Dennich, and T.Cole)
(2) July, 1984 Diploma (MSc-equivalent) in Biology (Microbiology, Botany, Organic Chemistry), University of Göttingen. Thesis: Experiments on 2D Crystallization and Electron Microscopical Analysis of the Purified Enzyme D- Ribulose-1,5-Bisphosphate Carboxylase. Supervisor: Prof. Dr. F. Mayer
(3) April, 1987 Doctor of Natural Sciences, University of Göttingen Thesis: Structure and Function-dependent Configurational Changes of D-Ribulose-1,5- Bisphosphate Carboxylase/Oxygenase from Alcaligenes eutrophus H16.
Supervisor: Prof. Dr. F. Mayer Co-referee: Prof. Dr. H.G. Schlegel
(4) July, 1987 Postdoctoral Fellow at the Biocentre of the University of Basel, Switzerland (Maurice E. Müller - Institute, Profs. Drs.U. Aebi & A. Engel)
(5) June, 1989 Guest Scientist (Institute for Microbiology, University of Göttingen, F.R.G., Prof. Dr. G. Gottschalk)
(6) Jan., 1990 Feodor-Lynen Research Associate at the University of Georgia, Athens, U.S.A. (Dept. of Biochemistry; Prof. Dr. L.G. Ljungdahl)
(7) Jan.,1991 Lecturer in Structural Molecular Biology (School of Biochemistry & Mol. Biology and School of Biology, University of Leeds, UK) with tenure from March 1994 onwards
(8) July,1999 and 2000 Visiting Professor, Institute of Virology, University of Marburg, F.R.G.
(9) Aug., 2000 Senior Lecturer (University of Leeds)
(10) Oct., 2000 Director of the Microscopy & Imaging Center & Professor of Biology (Texas A&M University, College Station, U.S.A.)
Recent Publications
Glykos, N.M., Holzenburg, A. and Phillips, S.E.V.: Low-resolution structural characterisation of the arginine repressor/ activator from Bacillus subtilis: A combined X-ray crystallographic and electron microscopical approach. Acta Cryst. D 54, 215-225 (1998).
Stoylova, S., Gray, E., Barrowcliffe, T.W., Kemball-Cook, G. and Holzenburg, A.: Structural determination of lipid-bound human blood coagulation factor IX.Biochim. Biophys. Acta 1383, 175-178 (1998).
Stoylova, S.S., Flint, T.D., Kitmitto, A., Ford, R.C and Holzenburg, A.: Comparison of photosystem II 3-D structure as determined by electron crystallography of frozen-hydrated and negatively stained specimens. Micron 29, 341-348 (1998).
Rishovd, S., Holzenburg, A., Johansen B.V. and Lindqvist, B.H.: Bacteriophage P2 and P4 Morphogenesis: Structure and function of the connector. Virology 245, 11-17 (1998).
Kitmitto, A., Mustafa, A.O., Holzenburg, A. and Ford, R.C.: Three-dimensional structure of higher plant photosystem I determined by electron crystallography. J. Biol. Chem. 273, 29592-29599 (1998).
Hoppert, M. and Holzenburg, A.: Electron Microscopy in Microbiology. RMS Handbook Series, Bios Scientific Publishers Ltd., Oxford (1998).
Stoylova, S.S., Ford, R.C. and Holzenburg, A.: Cryo-electron crystallography of small and mosaic 2-D crystals: An assessment of a procedure for high-resolution data retrieval. Ultramicroscopy 77, 113-128 (1999).
Pilling, A., Rosenberg, M.F., Willis, S.H., Jäger, J., Cohen, G.H., Eisenberg, R.J., Meredith, D.M. and Holzenburg, A.: The three-dimensional structure of herpes simplex virus type 1 glycoprotein D at 2.4 nm resolution. J. Virology 73, 7830-7834 (1999).
Kitmitto, A., Mustafa, A.O., Ford, J.W., Holzenburg, A. and Ford, R.C.: Does photoinhibition and/or phosphorylation of photosystem II influence its in vivo oligomeric state? Biochim. Biophys. Acta 1413, 21-30 (1999).
Stoylova, S.S., Lenting, P. J., Kemball-Cook, G. and Holzenburg, A.: Electron crystallography of human blood coagulation factor VIII bound to phospholipid monolayers. J. Biol. Chem. 274, 36573-36578 (1999).
Stoylova, S., Flint, T.D., Ford, R.C. and Holzenburg, A.: Structural analysis of 2-D crystals of photosystem II in far-red light adapted thylakoid membranes: New crystal forms provide evidence for a dynamic reorganization of light-harvesting antennae subunits. \ Eur. J. Biochem. 267, 1-10 (2000).
Simidjiev, I., Stoylova, S., Amenitsch, H., Javorfi, T., Mustardy, L., Laggner, P., Holzenburg, A. and Garab, G.: Self-assembly of large-ordered lamellae from non-bilayer lipids and integral membrane proteins in vitro. Proc. Natl. Acad. U.S.A 97, 1473-1476 (2000).
Xue, H., Zheng, H., Li, H.M., Kitmitto, A. Zhu, H., Lee, P. and Holzenburg, A.: A fragment of recombinant GABAA receptor alpha1 subunit forming rosette-like homo-oligomers. J. Mol. Biol. 296, 739-742 (2000).
Holzenburg, A. and Scrutton, N (eds.): Enzyme-catalysed electron and radical transfer. Subcellular Biochemistry Series. Plenum/Kluwer Publishers, New York (2000).
Ruffle, S.V., Mustafa, A.O., Kitmitto, A., Holzenburg, A.and Ford, R.C.: The location of the mobile electron carrier ferredoxin in vascular plant photosystem I. J. Biol. Chem. 275, 36250-36255 (2000).
Beales, L.P., Rowlands, D.J. and Holzenburg, A.: Structural motifs of the internal ribosome entry site (IRES) of hepatitis C virus visualised by electron microscopy. RNA 7, 661-670 (2001).
Rosenberg, M.F., Mao, Q., Holzenburg, A., Ford, R.C., Deeley, R.G. and Cole, P.C.: The structure of the multidrug resistance protein (MRP1/ABCC1). Crystallization and single particle analysis. J. Biol. Chem. 276, 16076-16082 (2001).
Scheffczik, H., Kraus, I., Kiermayer, S., Bogner, E., Holzenburg, A., Garten, W. and Eickmann, M.: Multimerization potential of the cytoplasmic domain of the human cytomegalovirus glycoprotein B. FEBS Lett. 506, 113-116 (2001).
Xue, H., Shi, H., Tsang, S.Y., Zheng, H., Savva, C.G.., Sun, J. and Holzenburg, A.: A recombinant glycine receptor fragment forms homo-oligomers distinct from its GABAA counterpart. J. Mol. Biol. 312, 915-920 (2001).
Holzenburg, A. and Bogner, E. (eds.): Structure-function relationships of human pathogenic viruses. Kluwer Academic/Plenum Publishers, New York (2002).
Holzenburg, A. and Bogner, E.: From concatemeric DNA into unitlength genomes - a miracle or clever genes ? In: Structure-function relationships of human pathogenic viruses (Holzenburg, A. and Bogner, E., eds.) pp. 155-170. Kluwer Academic/Plenum Publishers, New York (2002).
Ford, R.C., Stoylova, S.S. and Holzenburg, A.: An alternative model for photosystem II/light harvesting complex II in grana membranes based on cryo-electron microscopy studies. Eur. J. Biochem. 269, 326-336 (2002).
Stoilova-McPhie, S., Villoutreix, B.O., Mertens, K., Kemball-Cook, G. and Holzenburg, A.: Three-dimensional structure of membrane-bound coagulation factor VIII. Blood 99, 1215-1223 (2002).
Scheffczik, H., Savva, C., Holzenburg, A., Kolesnikova, L. and Bogner, E.: The terminase subunits pUL56 and pUL89 of human cytomegalovirus are DNA metabolizing proteins with toroidal structure. Nucleic Acid Research 30, 1695-1703 (2002).
Ruffle, S.V., Mustafa A.O., Kitmitto, A., Holzenburg, A. and Ford, R.C.: The location of plastocyanin in vascular plant photosystem I. J. Biol. Chem. 277, 25692-25696 (2002).
Garab, G., Cseh, Z., Kovacs, L., Rajagopal, S., Varkonyi, Z., Wentworth, M., Mustardy, L., Der, A., Ruban, A.V., Papp, E., Holzenburg, A. and Horton, P.: Light-induced trimer to monomer transition in the main light-harvesting antenna complex of plants: A thermo-optic mechanism. Biochemistry 41, 15121-15129 (2002).
Sharma, S., Arockiasamy, A., Ronning, D.R., Savva, C.G., Holzenburg, A., Braunstein, M., Jacobs, Jr., W.R. and Sacchettini, J.C.: Crystal structure of M. tuberculosis SecA, a preprotein translocating ATPase. Proc. Natl. Acad. Sci. U.S.A. 100, 2243-2248 (2003).
Beales, L.P., Holzenburg, A. and Rowlands, D.J.: Viral internal ribosome entry site structures segregate into two distinct morphologies. J. Virol. 77, 6574-6579 (2003).
Holzenburg, A.: Book review: Outline of Crystallography for Biologists by David Blow. Quart. Rev. Biol. 78, 221 (2003).
Hoppert, M. and Holzenburg, A.: Preparation Techniques. In: Michael Hoppert "Microscopic Techniques in Biotechnology" pp. 201-286. Wiley-VCH, Weinheim (2003).
Winslow, B.J., Cochran, M.D., Holzenburg, A., Sun, J., Junker, D.E. and Collisson, E.W.: Replication and expression of a swinepox virus vector delivering feline leukemia virus Gag and Env to cell lines of swine and feline origin. Virus Res. 98, 1-15 (2003).
Vakonakis, I., Sun, J., Wu, T., Holzenburg, A., Golden, S.S. and LiWang, A.C.: NMR structure of the KaiC-interacting C-terminal domain of KaiA, a circadian clock protein: implications for KaiA-KaiC interaction. Proc. Natl. Acad. Sci. U.S.A. 101, 1479-1484 (2004).
Deaton, J., Sun, J., Holzenburg, A., Struck, D.K., Berry, J. and Young, R.: Functional bacteriorhodopsin is efficiently solubilized and delivered to membranes by the chaperonin GroEL. Proc. Natl. Acad. Sci. U.S.A. 101, 2281-2286 (2004).
Savva, C.G., Holzenburg, A. and Bogner, E.: Insights into the structure of human cytomegalovirus large terminase subunit pUL56. FEBS Lett. 563, 135-140 (2004).
Deaton, J., Savva, C.G., Holzenburg, A., Berry, J. and Young, R.: Solubilization and delivery by GroEL of megadalton complexes of the lambda holin. Protein Sci. 13, 1778-1786 (2004).
Weiss, J.L., Evans, N.A., Ahmed, T., Wrigley, J.D.J., Khan, S., Wright,C., Keen, J.N., Holzenburg, A. and Findlay, J.B.C.: Methionine-rich repeat proteins: a family of membrane-associated proteins which contain unusual repeat regions. Biochim. Biophys. Acta 1668, 164-174 (2005).
Sun, J., Savva, C.G., Deaton, J., Kaback, R.H., Svrakic, M., Young, R. and Holzenburg, A.: Asymmetric binding of membrane proteins to GroEL. Arch. Biochem. Biophys. 434, 352-357 (2005).
Colangeli, R., Helb, D., Sridharan, S., Sun, J., Varma-Basil, M., Hazbón, M.H., Harbacheuski, R., Megjugorac, N.J., Jacobs Jr, W.R., Holzenburg, A., Sacchettini, J.C. and Alland, D.: The Mycobacterium tuberculosis iniA gene is essential for activity of an efflux pump that confers drug tolerance to both isoniazid and ethambutol. Mol. Microbiol. 55, 1829-1840 (2005).
Guarino, L.A., Bhardwaj, K., Dong.,W., Sun, J., Holzenburg, A. and Kao, C.: Mutational analysis of the SARS virus Nsp15 endoribonuclease: identification of residues affecting hexamer formation. J. Mol. Biol. 353, 1106-1117 (2005).
Colangeli, R., Helb, D., Sridharan, S., Sun, J., Varma-Basil, M., Hazbón, M.H., Harbacheuski, R., Megjugorac, N.J., Jacobs Jr, W.R., Holzenburg, A., Sacchettini, J.C. and Alland, D.: The Mycobacterium tuberculosis iniA gene is essential for activity of an efflux pump that confers drug tolerance to both isoniazid and ethambutol. Mol. Microbiol. 55, 1829-1840 (2005).
Sun, J., Savva, C.G., Deaton, J., Kaback, R.H., Svrakic, M., Young, R. and Holzenburg, A.: Asymmetric binding of membrane proteins to GroEL. Arch. Biochem. Biophys. 434, 352-357 (2005).
Weiss, J.L., Evans, N.A., Ahmed, T., Wrigley, J.D.J., Khan, S., Wright,C., Keen, J.N., Holzenburg, A. and Findlay, J.B.C.: Methionine-rich repeat proteins: a family of membrane-associated proteins which contain unusual repeat regions. Biochim. Biophys. Acta 1668, 164-174 (2005).
Bogner, E. and Holzenburg, A. (eds.): New concepts of antiviral therapy. Springer, Dordrecht (2006).
Hawkins, D.M., Trache A., Ellis, E.A., Stevenson D., Holzenburg, A., Meininger, G.A. and Reddy, S.M.: Quantification and confocal imaging of protein specific molecularly imprinted polymers. Biomacromolecules 7, 2560-2564 (2006).
Bhardwaj, K., Sun, J., Holzenburg, A., Guarino, L.A. and Kao, C.: RNA recognition and cleavage by the SARS coronavirus endoribonuclease. J. Mol. Biol. 361, 243-256 (2006).
Sun, J., Duffy, K.E., Ranjith-Kumar, C.T., Xiong, J., Lamb, R.J., Santos, J., Masarapu, H., Cunningham, M., Holzenburg, A., Sarisky, R.T., Mbow, M.L. and Kao, C.: Structural and functional analyses of the human Toll-like receptor 3. Role of glycosylation. J. Biol. Chem. 281, 11144-11151 (2006).
Hawkins, D.M., Ellis, E.A., Stevenson, D., Holzenburg, A. and Reddy, S.M.: Novel critical point drying (CPD) based preparation and transmission electron microscopy (TEM) imaging of protein specific molecularly imprinted polymers (HydroMIPs). J. Mater. Sci. 42, 9465-9468 (2007).
Heuck, A.P., Savva, C.G., Holzenburg, A. and Johnson, A.E.: Conformational changes that effect oligomerization and initiate pore formation are triggered throughout perfringolysin O upon binding to cholesterol. J. Biol. Chem. 282, 22629-22637 (2007).
Ranjith-Kumar, C.T., Miller, W., Sun, J., Xiong, J., Santos, J., Yarbrough, I., Lamb, R.J., Mills, J., Duffy, K.E., Hoose, S., Cunningham, M., Holzenburg, A., Mbow, M.L., Sarisky, R.T. and Kao, C.C.: Effects of single nucleotide polymorphisms on Toll-like receptor 3 activity and expression in cultured cells. J. Biol. Chem. 282, 17696-17705 (2007).
Ramesh, A., Savva, C.G., Holzenburg, A. and Sacchettini, J.C.: Crystal structure of Rsr, an ortholog of the antigenic Ro protein, links conformational flexibility to RNA binding activity. J. Biol. Chem. 282,14960-14967 (2007).
Sun, J., Dufort, C., Daniel, M.C., Murali, A., Chen, C., Gopinath, K., Stein, B., De, M., Rotello, V.M., Holzenburg, A., Kao, C.C. and Dragnea, B.: Core-controlled polymorphism in virus-like particles. Proc. Natl. Acad. Sci. U.S.A. 104, 1354-1359 (2007).
Smith A.G., Johnson, C.B., Ellis, E.A., Vitha, S. and Holzenburg, A: Protein screening using cold microwave technology. Anal. Biochem. 375, 313-317 (2008).
Ford, R.C. and Holzenburg, A.: Electron crystallography of biomolecules: mysterious membranes and missing cones. Trends Biochem. Sci. 33, 38-43 (2008).
