Research Interests

The Robinson laboratory is interested in understanding the fundamental interactions between molecules, both in isolation and in the complex environment of the cell. We use our growing understanding to design proteins with more robust or novel properties and to engineer cellular systems for improved production or drug screening applications. To this end, we are investigating the determinants of protein folding and misfolding, on both atomic and molecular levels. We have developed several novel approaches to inhibit protein misfolding and aggregation. Additionally, we are designing cellular systems for optimal expression of membrane proteins and antibodies.

A major goal of this research is to establish a set of cellular systems that could express any protein of interest. Recently, we have begun studies to understand the cellular interactions that lead to hyperphosphorylation and aggregation of tau protein, which is relevant to Alzheimer’s disease, and several other neurodegenerative diseases including corticobasal degeneration.

Our approach uses techniques in molecular biology, genetic engineering, and biophysical chemistry to identify and study macromolecules at both an atomic and cellular level. Mechanistic modeling guides us in experimental design and analysis. We use both prokaryotic (bacterial) systems as well as eukaryotic systems, such as yeast or mammalian cells. The research in the laboratory has focus areas in protein stability, expression, and aggregation for biotechnology and biomedical applications.

Education

B.S., Johns Hopkins University, 1988
M.S., Johns Hopkins University, 1989
Ph.D., University of Illinois at Urbana-Champaign, 1994

Biographical Sketch

Anne Skaja Robinson joined Tulane University in January 2012 as the Chair of Chemical and Biomolecular Engineering. Prior to Tulane, Dr. Robinson was a Full Professor and Associate Chair at the University of Delaware, where she started her academic career. She earned her Bachelor’s and Master’s degrees in Chemical Engineering from the Johns Hopkins University. She earned her PhD in Chemical Engineering from the University of Illinois at Urbana-Champaign, and was an NIH postdoctoral fellow in the Department of Biology at MIT before joining the faculty at UD. Her honors include a DuPont Young Professor Award, and a National Science Foundation Presidential Early Career Award for Science and Engineering (PECASE) Award, and more recently being inducted into AIMBE.

Teaching

• Biomolecular and Cellular Engineering

Selected Publications

Mass and Heat Transfer Textbook

1. Russell, T.W.F., Robinson, A.S., and Wagner, N.J., (2008) Mass and Heat Transfer: Analysis of Mass Contactors and Heat Exchangers, Cambridge University Press, Cambridge, UK (www.cambridge.org/9780521886703).

G-protein Coupled Receptors (7-Helix Transmembrane Proteins)

1. Robinson, A.S., ed (2011) Production of Membrane Proteins – Strategies for Expression and Isolation, Wiley VCH.
2. O’Malley, M.A., Naranjo, A.N., Lazarova, T., Robinson, A.S.* (2010) “Analysis of Adenosine A2a Receptor Stability: Effects of Ligands and Disulfide Bonds,” Biochemistry, Nov 2;49(43):9181-9.
3. O'Malley, M.A., Helgeson, M.E., Wagner, N.J., Robinson, A.S.* (2011) The morphology and composition of cholesterol-rich micellar nanostructures determine transmembrane protein (GPCR) Activity, Biophysical Journal, 100(2): L11-13.
4. Robinson, A.* (2011) “New Tools for Breaking Barriers to GPCR Expression in E. coli” Journal of Molecular Biology. 408 (4): 597-598.
5. Britton, Z., Young, C.L., Can, O., McNeely,P., Naranjo, A.N., Robinson, A. S.* (2011), “Membrane protein expression in yeast Saccharomyces cerevisiae” in Production of Membrane Proteins – Strategies for Expression and Isolation, Wiley VCH, Robinson, A.S., ed.
6. O'Malley MA, Mancini JD†, Young CL, McCusker EC, Raden D, Robinson A.S*. (2009) “Progress toward heterologous expression of active G-protein-coupled receptors in Saccharomyces cerevisiae: Linking cellular stress response with translocation and trafficking.” Protein Sci. 18(11):2356-70.
7. McCusker, E., and Robinson, A.S.*, (2008) "Refolding of G protein α subunits from inclusion bodies expressed in Escherichia coli," Protein Exp. Purif., Apr;58(2): 342-55. Epub 2007 Dec 8.
8. McCusker, E., Bane, S.E., O’Malley, M., and Robinson, A.S.* (2007), “Heterologous GPCR expression: A bottleneck to obtaining crystal structures”, Biotech Progress, May-Jun;23(3):540-7.
9. O’Malley, M., Lazarova, T., Britton, Z.T., and Robinson, A.S. * (2007) “High-level expression in Saccharomyces cerevisiae enables isolation and spectroscopic characterization of functional human adenosine A2a receptor”, J. Struct Biol., 159(2): 166-178.
10. Bane, S.E., Velasquez, J.E. †, and Robinson, A.S. * (2007) “Expression and purification of milligram levels of inactive G-protein coupled receptors in E. coli”, Protein Expression and Purification, 52(2):348:355.
11. Niebauer, R. T., and Robinson, A.S. * (2006) “Exceptional total and functional yields of the human adenosine (A2a) receptor expressed in the yeast Saccharomyces cerevisiae”, Prot. Exp. Purif., 46, p. 204-211.
12. Wedekind, A.L. †, O’Malley, M., Niebauer, R.T., and Robinson, A.S. * (2006) Optimization of the Human Adenosine A2a Receptor Yields in Saccharomyces cerevisiae, Biotechnology Progress, 22(5):1249-55.
13. Niebauer, R. T. and Robinson, A.S.(2004) “Saccharomyces cerevisiae protein expression: From protein production to protein engineering” in Expression Technologies, Horizon Scientific Press.
14. Niebauer, R.T., Wedekind, A. and Robinson, A.S. * (2004) “Decreases in yeast expression yields of the human adenosine receptor are a result of translational or post-translational events”, Protein Exp. Purif., 37 (1) 134-143.
15. Butz, J., Niebauer, R. T., and Robinson, A.S. (2003), “Co-expression of molecular chaperones does not improve the heterologous expression of mammalian G-Protein coupled receptor expression in yeast,” Biotech. Bioeng, 84 (3) 292-304.

Stress Response And Chaperone Interactions

1. Young C.L., Yuraszeck T., Robinson A.S.* (2011) “Decreased secretion and unfolded protein response upregulation,” Methods Enzymol. 491:235-60.
2. Griesemer, M., Young, C.L., Doyle III, F.J., Robinson, A.S. and Petzold, L.* (2011) Spatial Localization of Chaperone Distribution in the Endoplasmic Reticulum of Yeast, IET Systems Biology, in press.
3. Spatara, ML and Robinson, A.S.* (2010) “Transgenic mouse and cell culture models demonstrate a lack of mechanistic connection between endoplasmic reticulum stress and tau dysfunction” Journal of Neuroscience Research, 88(9):1951-61.
4. Yuraszeck, T.M., Neveu, P., Rodriguez-Fernandez, M., Robinson, A.S., Kosik, K.S., Doyle III, F.J.* (2010) "Vulnerabilities in the Tau Network and The Role of Ultrasensitive Points in Tau Pathophysiology", PLoS Computational Biology, 6(11): e1000997. doi:10.1371/journal.pcbi.1000997 (with cover art).
5. Xu, P. and Robinson, A.S.* (2009) “Decreased secretion and unfolded protein response up-regulation are correlated with intracellular retention for single-chain antibody variants produced in yeast” Biotech & Bioeng, 104(1):20-9.
6. Hildebrandt, S.,Raden, D, Petzold, L, Robinson, A.S., and Doyle III, F.J.* (2008) “A top-down approach to mechanistic biological modeling: application to the single-chain antibody folding pathway”, Biophysical Journal, 95(8):3535-58. Epub 2008 Jul 18.
7. Famá, M.C., Raden, D., Zacchi, N., Lemos, D.R., Robinson, A.S., and Silberstein, S. * (2007) “The Saccharomyces cerevisiae YFR041C/ERJ5 gene encoding a type I membrane protein with a J domain is required to preserve the folding capacity of the endoplasmic reticulum” Biochim Biophys Acta, 1773(2):232-42.
8. Griesemer, M., Young, C., Raden, D., Petzold, L., Robinson, A.S., Doyle, F.J. * (2007) “Computational Modeling of  Chaperone Interactions in the Endoplasmic Reticulum of Saccharomyces cerevisiae.” Proc. Int. Conf. Foundations of Systems Biology, Stuttgart, Germany.
9. Yuraszeck, T., Raden, D, Robinson, A.S., and Doyle, F.J.* (2007) “Microarray Analysis of the Unfolded Protein Response in S. cerevisiae Reveals Evidence of Down-regulation.” Proc. Int. Conf. Foundations of Systems Biology, Stuttgart, Germany.
10. Xu, P., Raden, D., Doyle, F.J. III, and Robinson, A.S. * (2005) “Analysis of unfolded protein response during single-chain antibody expression in Saccaromyces cerevisiae reveals different roles for BiP and PDI in folding”, Metabolic Engineering, 7 (4) 269-279.
11. Raden, D., Hildebrandt, S, Xu, P., Bell, E. †, Doyle, III, F.J. and Robinson, A.S. * (2005), “Analysis of cellular response to protein overexpression.” IEE Proceedings: Systems Biology 152 (4) 285-289.
12. Hildebrandt, S., D. Raden, E. Bell†, Robinson, A.S. , and F.J. Doyle III* (2005) “Modeling the Unfolded Protein Response in Saccharomyces Cerevisiae”, Proc. Int. Conf. Foundations of Systems Biology, Santa Barbara, California.
13. Kauffman, K., Pridgen, E.M., Doyle, F.J. III, Dhurjati, P., and Robinson, A.S. (2002) “Decreased Protein Expression and Oscillating BiP Levels Result during Heterologous Protein Expression in S. cerevisiae,” Biotech. Prog., 18, 942-940. DOI: 10.1021/bp025518g
14. Kauffman, K., Dhurjati, P. , Robinson, A.S. and F.J. Doyle III , “Framework for Modeling Information Flow in Biological Processes: Application to the Unfolded Protein Response.” Proc. IFAC Conf. Comput. Appl. Biotech (CAB), 2001.
15. Robinson, A.S., Bockhaus, J.A., Voegler, A.C.and Wittrup, K.D. (1996) “Reduction of BiP levels decreases heterologous protein secretion in Saccharomyces cerevisiae” J. Biol. Chem. 271, 10017-10022.
16. Robinson, A.S. and Lauffenburger, D.A.(1996) “Model for ER Chaperone Dynamics and Secretory Protein Interactions.” AIChE J. 42, 1443-1453.
17. Robinson, A.S. and K.D. Wittrup (1995) “Constitutive Overexpression of Secreted Heterologous Proteins Decreases Extractable BiP and PDI Levels in Saccharomyces cerevisiae.” Biotech Prog.  11, 171-177.
18. Wittrup , K.D., Robinson, A.S., Parekh, R.N. and Forrester, K.J. (1994) “Existence of an Optimal Expression Level for Secretion of Foreign Proteins in Yeast.” Ann. N.Y. Acad. Sci.  745, 321-330.
19. Robinson, A.S., V. Hines, and K.D. Wittrup (1994) “Overexpression of Protein Disulfide Isomerase Increases Secretion of Foreign Proteins in the Yeast Saccharomyces cerevisiae.” Bio/Tech. 12, 381-384.
20. "Foreign Proteins in Eucaryotic Cells.”  in Protein Folding:  In vivo and In vitro.  ACS Symposium Series 526.  Jeffrey Cleland, Ed., 121-132.
21. Robinson, A.S. and Wittrup, K.D. . “Methods for Increasing Secretion of Overexpressed Proteins.” Patent # 5,773,245.  Filed 10/92.  Accepted 6/30/98.

Protein Aggregation and Refolding

1. Sahin, E., Jordan, J.L., Spatara, M.L., Naranjo, A.N., Weiss IV, W.F., Robinson, A.S, Fernandez, E.J.*, Roberts, C.J.* (2011) “Computational Design and Biophysical Characterization of Aggregation-Resistant Point Mutations for γD Crystallin Illustrate a Balance of Conformational Stability and Intrinsic Aggregation Propensity”, Biochemistry, Feb 8;50(5):628-39.
2. Spatara, ML, Roberts, CJ, and Robinson, A.S.* (2009) “Kinetic folding studies of the P22 tailspike beta-helix domain reveal multiple unfolded states.” Biophys Chem. 141(2-3):214-21.
3. Webber T, Gurung S, Saul J, Baker T, Spatara M, Freyer M, Robinson A.S., and Gage MJ* (2009) “The C-terminus of the P22 tailspike protein acts as an independent oligomerization domain for monomeric proteins.”, Biochem J. 2009 May 1;419(3):595-602.
4. Gage, M.J, Lefebvre, B.G., and Robinson, A.S.* (2006) “Determinants of Protein Folding and Aggregation in P22 Tailspike,” in Misbehaving Proteins, ACS Publications, eds. Regina Murphy and Amos Tsai.
5. Kim, J. and Robinson, A.S. * (2006) Dissociation of intermolecular disulfide bonds in P22 tailspike protein intermediates in the presence of SDS, Protein Science, 15 (7), p. 1791-3.
6. Gage, M.J., Zak, J. and Robinson, A.S. * (2005) “Three Amino Acids that are Critical to Formation and Stability of the P22 Tailspike Trimer”, Protein Science, 14 (9) 2333-43.
7. Lefebvre, B.G., Gage, M.J., and Robinson, A.S. (2004) “Maximizing Recovery of Native Protein from Aggregates by Optimizing Pressure Treatment,” Biotechnology Progress, 20, 2, p. 623-629.  10.1021/bp034221v.
8. Lefebvre, B.G., Comolli, N.K., Gage, M.J. and A.S. Robinson * (2004), “Pressure dissociation studies provide insight into oligomerization competence of temperature-sensitive mutants of P22 tailspike,” Protein Sci., 13 (6) 1538-46.
9. Danek, B.L. and Robinson, A.S.* (2004) “P22 tailspike trimer assembly is governed by interchain redox associations,” Biochem. Biophys. Acta, 1700(1):105-16. [5]
10. Gage, M.J. and Robinson, A. S. (2003) “C-terminal Hydrophobic Interactions Play a Critical role in Oligomeric Assembly of the P22 Tailspike Trimer,” Protein Sci., 12, 12, p. 2732-47.
11. Lefebvre, B.G., and Robinson, A.S. (2003), “Pressure treatment of tailspike aggregates rapidly produces on-pathway folding intermediates,” Biotech. Bioeng, 82, 5, p. 595-604. DOI: 10.1002/bit.10607
12. Danek, B.L., and Robinson, A. S. (2003) “Non-native interactions between cysteines direct productive assembly of P22 tailspike protein,” Biophys J., 85, 5, p. 1-11.
13. Sinacola, J. and Robinson, A. S. (2002) “Rapid refolding and polishing of single-chain antibodies from E. coli inclusion bodies” Protein Exp. Purif., Vol. 26, No. 2, Nov 2002, pp. 301-308. DOI: 10.1016/S1046-5928(02)00538-7.
14. Haase-Pettingell, C., Betts, S., Raso, S.W., Stuart, L., Robinson, A.S. and J. King (2001), “Role for Cysteine Residues in the In Vivo Folding and Assembly of the Phage P22 Tailspike,” Protein Sci. 10, 397-410.
15. Robinson, A.S. and J. King , (1997) “Disulfide-Bonded Intermediate on the Folding and Assembly Pathway of a Non-Disulfide Bonded Protein.” Nature Struct. Biol., 4, 450-455.
16. Foguel, D., Robinson, C.R., Caetano de Sousa Jr., P., Silva, J. L., Robinson A.S. (1999), “Hydrostatic Pressure Rescues Protein Aggregates”, Biotech. Bioeng. 63, 552-558.
17. Robinson, A.S., D. Foguel, J.L. Silva, C.R. Robinson. “Use of Hydrostatic Pressure to Inhibit and Reverse Protein Aggregation and Facilitate Protein Refolding.” US Patent # 7,615,617. Filed 10/99. Issued 11/10/09.
18. King , J., Haase-Pettingell, C., Robinson, A. S., Speed, M and Mitraki, A. (1996) “Thermolabile Folding Intermediates: Inclusion Body Precursors and Chaperonin Substrates” FASEB J., 10, 57-66.

Extermophilic Protein Expression – beta Glucosidase (CelB) and Pyrolysin

1. Powers, S.L. and Robinson, A. S.* (2007) “PDI Improves Secretion of Redox-Inactive b-glucosidase”, Biotech Prog., Mar-Apr;23(2):364-9. E-pub Feb 22, DOI: 10.1021/bp060287p
2. Powers, S.L., Robinson, C.R., and Robinson, A.S. * (2007) Denaturation of an Extremely Stable Hyperthermophilic Protein Occurs via a Dimeric Intermediate, Extremophiles, 11(1):179-89.
3. Smith, J.D., Richardson, N.E. and Robinson, A. S.* (2005) “Elevated expression temperature in a mesophilic host results in increased secretion of a hyperthermophilic enzyme and decreased cell stress,” Biochem. Biophys. Acta, 1752 (1) 18-25.
4. Smith, J.D., Tang, B.C., and Robinson, A. S.(2004) “Protein disulfide isomerase, but not binding protein, overexpression enhances secretion of a non-disulfide-bonded protein in yeast”, Biotech. Bioeng., 85, 3, p. 340-50.
5. Smith, J.D. and Robinson, A. S. (2002) “Expression of an archael enzyme in a eucaryotic host: A secretion bottleneck at the ER,” Biotech. Bioeng., 79, 7, p. 713-723. DOI: 10.1002/bit.10367
6. Blumentals, I. I., Robinson, A. S. and Kelly, R. M.. (1990). "Characterization of Sodium Dodecyl Sulfate Resistant Proteolytic Activity in the Hyperthermophilic Archaebacterium Pyrococcus furiosus." Appl. Envir. Microbiol. 56, 1992-1998.
7. Blumentals, I.I., S.H. Brown, R.N. Schicho, A.K. Skaja [Robinson], H.R. Costantino, and R.M. Kelly . (1990) “The Hyperthermophilic Archaebacterium, Pyrococcus furiosus:  Development of Culturing Protocols, Perspectives on Scale-Up, and Potential Applications.” Ann. N.Y. Acad. Sci., 589, 301-314.
8. Kelly, R.M., Robinson,A.K.S., I.I. Blumentals, S.H. Brown, and C.B. Anfinsen.  “Proteolytic Enzymes from Hyperthermophilic Bacteria and Processes for Their Production.” 
   Patent # 5,242,817.  Filed 9/12/89.  Accepted 9/7/93.  Licensed to Takara Shuzo.

Other

1. Blumentals, I. I., Kelly, R. M., A. K. Skaja [Robinson] and J. Shiloach. (1987) "Effect of Culturing Conditions on the Production of Exotoxin A by Pseudomonas aeruginosa."  Ann N Y Acad. Sci. 506, 663-668.
2. Forsten-Williams*, K.F., Cassino, T.R, Delo, L.J., Bellis, A.D., Robinson, A.S., and Ryan, T.E., (2007) Enhanced Insulin-like Growth Factor-I (IGF-I) Cell Association at Reduced pH is Dependent on IGF Binding Protein-3 (IGFBP-3) Interaction, Journal of Cellular Physiology, 210(2):298-308.