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Robert L. Campbell PhD
 Robert L. Campbell
Position(s)
Assistant Professor
Contact Info
Botterell Hall, Room 644
613-533-6821

Faculty Bio

My research in collaboration with Dr. Peter Davies' lab has involved the use of X-ray crystallography and molecular modelling to study both the calpain family of enzymes and ice-binding proteins. 

Calpains are a family of cysteine proteases that depend on the presence of Ca2+ to allow them to adopt an active conformation. We are studying their structures to try to understand the details of this activation process. We are also using X-ray crystallography and modelling to determine structures of the enyzme in combination with substrates and inhibitors for use in structure-based inhibitor design. Specific inhibitors may be useful in further research studies and as potential lead compounds for the development of pharmaceuticals. 

Ice-binding proteins (IBPs) can be divided into two main groups: antifreeze proteins (AFPs) that inhibit ice crystal growth and ice nucleating proteins (INPs) that stimulate ice crystal growth.  AFPs bind to the surfaces of ice crystals and inhibit their growth at subzero temperatures, thereby protecting the host organism from freezing damage. We have hypothesized that AFPs bind to ice by first ordering water molecules at the protein's surface to be in an ice-like array. In order to explore this idea, we are using X-ray crystallography combined with homology modelling, molecular dynamics and site-directed mutagenesis to examine the ordering of water molecules around the proteins' surfaces. INPs appear to differ from AFPs predominantly by the size of their ice-binding site.  We are attempting to engineer INPs from AFPs and vice versa.

Research Interests

My research in collaboration with Dr. Peter Davies' lab has involved the use of X-ray crystallography and molecular modelling to study both the calpain family of enzymes and ice-binding proteins.

Calpains are a family of cysteine proteases that depend on the presence of Ca2+ to allow them to adopt an active conformation. We are studying their structures to try to understand the details of this activation process. We use X-ray crystallography and modelling to determine structures of the enyzme in combination with substrates and inhibitors for use in structure-based inhibitor design. Specific inhibitors may be useful in further research studies and as potential lead compounds for the development of pharmaceuticals for the treatment after ischemic reperfusion events.

Antifreeze proteins (AFPs) are one class of ice-binding proteins that bind to the surfaces of ice crystals and inhibit their growth at subzero temperatures, thereby protecting the organism from freezing damage. We have hypothesized that AFPs bind to ice by first ordering water molecules at the protein's surface to be in an ice-like array. In order to explore this idea, we are using X-ray crystallography and homology modelling along with site-directed mutagenesis to examine the ordering of water molecules around the proteins' surfaces.  Another class of ice-binding proteins are those that nucleate the formation of ice (ice-nucleating proteins, INPs) that appear to differ from AFPs predominantly by the size of their ice-binding site.  We are attempting to explore this relationship by engineering INPs from AFPs and vice versa.

Selected Publications

  1. McCartney CE, Ye Q, Campbell RL, Davies PL. “Insertion sequence 1 from calpain-3 is functional in calpain-2 as an internal propeptide.” (2018) J Biol Chem. [Epub ahead of print] PubMed: 30254072
  2. Ye Q, Campbell RL, Davies PL. “Structures of human calpain-3 protease core with and without bound inhibitor reveal mechanisms of calpain activation.” (2018) J Biol Chem. 293, 4056-4070 PubMed: 29382717
  3. Guo S, Stevens CA, Vance TDR, Olijve LLC, Graham LA, Campbell RL, Yazdi SR, Escobedo C, Bar-Dolev M, Yashunsky V, Braslavsky I, Langelaan DN, Smith SP, Allingham JS, Voets IK, Davies PL. Structure of a 1.5-MDa adhesin that binds its Antarctic bacterium to diatoms and ice. (2017) Sci Adv. 3, e1701440 PubMed: 28808685
  4. Stevens CA, Semrau J, Chiriac D, Litschko M, Campbell RL, Langelaan DN, Smith SP, Davies PL, Allingham JS. “Peptide backbone circularization enhances antifreeze protein thermostability.” (2017) Protein Sci. 26, 1932-1941. PubMed: 28691252
  5. Low KE, Ler S, Chen K, Campbell RL, Hickey JL, Tan J, Scully CC, Davies PL, Yudin AK, Zaretsky S. “Rational design of calpain inhibitors based on calpastatin peptidomimetics.” (2016) J. Med. Chem.59, 5403-15 PubMed: 27148623
  6. Jung W, Campbell RL, Gwak Y, Kim JI, Davies PL, Jin E. “New Cysteine-Rich Ice-Binding Protein Secreted from Antarctic Microalga, Chloromonas sp.” (2016) PLoS One11, e0154056 PubMed: 27097164
  7. Basu K, Campbell RL, Guo S, Sun T, Davies PL “Modeling repetitive, non-globular proteins” (2016) Protein Sci25, 946-58 PubMed: 26914323.
  8. Sun T, Gauthier SY, Campbell RL, Davies PL (2015) Revealing Surface Waters on an Antifreeze Protein by Fusion Protein Crystallography Combined with Molecular Dynamic Simulations. J Phys Chem B. 119.12808-15. PubMed 26371748 (doi: 10.1021/acs.jpcb.5b06474)
  9. Basu, K, Graham, LA, Campbell, RL and Davies, PL (2015) Flies expand the repertoire of protein structures that bind ice. Proc Natl Acad Sci U S A.  112(3), 737-42. PubMed: 25561557 (doi: 10.1073/pnas.1422272112.)
  10. Low, KE, Partha, SK, Davies, PL, Campbell, RL (2014) Allosteric inhibitors of calpains: Reevaluating inhibition by PD150606 and LSEAL. Biochim Biophys Acta. 1840(12), 3367-3373.PubMed: 25196359
  11. Vance, TDR, Olijve, LLC, Campbell, RL, Voets, IK, Davies, PL, Guo, S, (2014) Ca2+-stabilized adhesin helps an Antarctic bacterium reach out and bind ice. Bioscience Reports. 34(4). PubMed: 24892750
  12. Partha, SK, Ravulapalli, R, Allingham, JS, Campbell, RL, Davies, PL (2014) Crystal structure of calpain-3 penta-EF-hand domain: A homodimerized PEF family member with calcium bound at the fifth EF-hand. FEBS Journal. 281(14), 3138-49. PubMed: 24846670
  13. Sun,T, Lin, F-H, Campbell, RL, Allingham, JS, and Davies, PL (2014) An antifreeze protein folds with an interior network of over 400 semi-clathrate waters. Science 343(6172), 795-8. PubMed: 24531972
  14. Guo, S, Garnham, CP, Partha, SK, Campbell, RL, Allingham, JS, Davies, PL (2013) Role of Ca2+ in folding the tandem β-sandwich extender domains of a bacterial ice-binding adhesion. FEBS Journal Ref. no.: FJ-13-0467.R1.PubMed: 24024640
  15. Campbell RL, Davies PL (2012) Structure-function relationships in calpains. Biochem J. 447, 335-51 PubMed: 23035980
  16. Middleton, AJ, Marshall, CB, Faucher, F, Bar-Dolev, M, Braslavsky, I, Campbell, RL, Walker, VK and Davies, PL (2012) Antifreeze protein from freeze-tolerant grass has a beta-roll fold with an irregularly structured ice-binding site. J. Mol. Biol. 416, 713-724 PubMed: 22306740
  17. Garnham, CP, Campbell, RL, Walker, VK and Davies, PL (2011) Novel dimeric beta-helical model of an ice nucleation protein with bridged active sites. BMC Structural Biology. 11:36  PubMed: 21951648
  18. Garnham, CP, Campbell, RL and Davies, PL (2011) Anchored clathrate waters bind antifreeze proteins to ice. Proc Natl Acad Sci 108, 7363-7367.  PubMed: 21482800 PubMed Central: PMC3088597
  19. Kelly, JC, Cuerrier, D, Graham, LA, Campbell, RL, and Davies, PL (2009) Profiling of calpain activity with a series of FRET-based substrates. Biochim Biophys Acta. 1794, 1505-1509. PubMed: 19555780
  20. Garnham, CP, Hanna, RA, Chou, JS, Low, KE, Gourlay, K, Campbell, RL, Beckmann, JS and Davies, PL (2009) Limb-girdle muscular dystrophy type 2A can result from accelerated autoproteolytic inactivation of calpain 3. Biochemistry 48, 3457-3467.  PubMed: 19226146
  21. Ravulapalli, R, Campbell, RL, Gauthier, SY, Dhe-Paganon, S and Davies PL (2009) Distinguishing between calpain heterodimerization and homodimerization. FEBS J. 276, 973-982.  PubMed: 19215300
  22. Hanna, RA, Campbell, RL and Davies, PL (2008) Calcium-bound structure of calpain and its mechanism of inhibition by calpastatin. Nature 456, 409-412.  PubMed: 19020623
  23. Qian, J, Cuerrier, D, Davies, PL, Li, Z, Powers, JC and Campbell, RL (2008) Cocrystal structures of primed side-extending alpha-ketoamide inhibitors reveal novel calpain-inhibitor aromatic interactions. J. Med. Chem. 51, 5264-5270. PubMed: 18702462
  24. Graham, LA, Marshall, CB, Lin, F-H, Campbell, RL and Davies, PL (2008) Hyperactive antifreeze protein from fish contains multiple ice-binding sites. Biochemistry 47, 2051-2063. PubMed: 18225917
  25. Garnham, CP, Gilbert, JA, Hartman, CP, Campbell, RL, Laybourn-Parry, J and Davies PL (2008) A Ca2+-dependent bacterial antifreeze protein domain has a novel beta-helical ice-binding fold. Biochem J. 411, 171-180.  PubMed: 8095937
  26. Cuerrier, D, Moldoveanu, T, Campbell, RL, Kelly, J, Yoruk, B, Verhelst, SHL, Greenbaum, D, Bogyo, M and Davies, PL (2007) Development of calpain-specific inactivators by screening of positional-scanning epoxide libraries. J. Biol. Chem. 282, 9600-9611.  PubMed: 17218315
  27. Cuerrier, D, Moldoveanu, T, Inoue, J, Davies, PL, Campbell RL (2006) Calpain Inhibition by Ketoamide and Cyclic Hemiacetal Inhibitors Revealed by X-ray Crystallography Biochemistry, 45, 7446-7452.  PubMed: 16768440

Campbell, Robert Lab

See my personal web page for more information.