Welcome to
PERIYANNAN RESEARCH GROUP
      line   
eiulogo

Home Gopal R Periyannan Research Teaching



                                Research

                                Group Members

                                Publications
                               
                                Protease Links

                                Other Links

                                Protein Instability Index Calculator
                              
                              
Research
One third of all known proteins are metalloproteins. The number of important enzymes containing metal ions in their active site is increasing; which thereby underscores the significance of the role played by metalloenzymes in biology and medicine. Our research focuses on the structure and the mechanistic aspects of Zn-metalloproteases in order to understand their physiological function and their role in disease development. 

Zinc Metalloproteases


Glutamate Carboxypeptidase II (GCPII)  

GCPII is a 750 residue, dimeric, transmembrane, Zn-metalloprotease expressed in the central nervous system and other types of tissues. The exact roles played by the members of GCPII class of proteins  in different types of tissues, other than neuronal tissues, are not well understood. GCPII is expressed in large amounts in prostate cancer tissue and in the newly forming blood vessels of most solid tumors hence thought to be contributor of cancer progression. In addition to the peptidase activity, GCPII is predicted to have cell signaling properties. Although considerable structural characterization has been carried out on GCPII, including crystal structures of the extracellular domains, the mechanism of catalysis remains unproved. A precise understanding of the steps involved in the catalytic mechanism of NAAG hydrolysis will immensely increase the scope of synthesizing novel drugs, including candidates that can cross the blood-brain barrier. Further, it is essential to probe for any signaling event associated with substrate binding or catalysis to reveal any other roles of GCPII. Such studies are hampered by the absence full-length GCPII or a homolog. Currently available GCPII expression systems provide only the Extracellular portion of the protein, and that in small quantity and at a great expense. We propose to overcome this hurdle by studying a suitable model protein for GCPII in an appropriate model organism.

 

cbriggs  GCPII Model Protease from Caenorhabditis elegans

We explored the genomic databases and identified a transmembrane Zn metalloprotease in nematode C. elegans with remarkable structural homology to GCPII, including the M28 family of Zn protease domain, transferring receptor like dimerization domain and potential for glycosylation. These structural similarities between the metalloprotease from S. pombe and GCPII prompt us to propose this putative protein as a suitable model to study structure-function relationships and the hydrolytic mechanism of GCPII class of proteins. We use spectroscopic techniques to probe the Zn(II) mediated catalytic mechanism of GCPII.


CauC_Single  Metalloprotease from Caulobacter crescentus

As a part of our investigation of Zn-metalloproteases and their roles in disease development, we have cloned and successfully tested the expression of a Zn-metalloprotease from Caulobacter crescentus, a gram negative bacterium, which is an important model organism for study of the cell cycle and cellular differentiation. The cloned metalloprotease shares amino acid sequence similarity with GCPII. Information gathered on the active site and the catalytic mechanism of this metalloprotease is expected to help understand the catalytic mechanism of GCPII.

                                                          

Biofuel

Microbial Biomass Conversion
Biofuels hold the promise to supplement our future energy needs in a renewable and sustainable manner. Recently there have been reports of conversion of plant-derived biomass into fuel quality carbon molecules through the combination of biological and chemical means. The feedstock molecules for these conversion processes have been mostly monosaccharides derived from plant polysaccharides. There is a continued search for an efficient enzyme system which could breakdown the unusual plant polysaccharides such as cellulose, pectin and xylan in a cost effective manner into their sugar makeup.

More updates on will be posted on our study of microbial polysaccharide metabolising enzymes as it uncovers.

           Go to Top                
Group Members
Principal Investigator
Gopal R Periyannan

Graduate
Hashni Epa Vidanage

Undergraduate
Current
Xa Burton
Andrew Hladilek

Alumni
Graduate
Dilani G Gamage - PhD - Wayne State University, Detroit, MI; Postdoctoral Fellow - Cicinnati Children's Hospital

Undergraduate
Andrew Bolokowicz
Brant Riegel
Breanne Cornwell
Driscolle Augustine
Jason DeGroate
John Saathoff
Kevin Vessells
Nicholas Kucinski
Timothy Russell
Scott Keller
Valencia Anderson

                       
Publications

1. Hu, Z., Periyannan, G., Bennett, B., and Crowder, M.W., “Role of the Zn1 and Zn2 sites in metallo-β-lactamase L1” J. Am. Chem. Soc. (2008) Oct 3 (Epub).

2. Z. Hu, G.R. Periyannan, M.W. Crowder, "Folding strategy to prepare Co(II)-substituted metallo-beta-lactamase L1"  Anal Biochem. (2008) Jul 15;378(2):177-83.

3. Amit Kumar, Gopal R. Periyannan, Beena Narayanan, Aaron W. Kittell,  Jung-Ja Kim and Brian Bennett. Experimental evidence for a metallohydrolase mechanism in which the nucleophile is not delivered by a metal ion: EPR spectrokinetic and structural studies of aminopeptidase from Vibrio proteolyticus. Biochem J. (2007) May 1;403(3):527-36. 

4. M.L. Matthews, G. R. Periyannan, T.K. Sigdel, C. Hajdin, B. Bennett, and M.W. Crowder, “Probing the reaction mechanism of the D-ala-D-ala dipeptidase, VanX, by using stopped-flow kinetic and rapid-freeze quench EPR studies on the Co(II)-substituted enzyme” J. Am. Chem. Soc. (2006) Oct 11;128(40):13050-
1.

5. A. L. Costello, G. R. Periyannan, K.-W. Yang, M. W. Crowder and D. L. Tierney, "Site Selective Binding of Zn(II) to Metallo-b-Lactamase L1 from Stenotrophomonas maltophilia", (2006) JBIC,  Apr;11(3):351-8.

6. Gopal R. Periyannan, A.L. Costello, D.L. Tierney, K.W. Yang, B. Bennett, and M.W. Crowder,  "Sequential Binding of Co(II) to Metallo-b-Lactamase CcrA" Biochemistry (2006) Jan 31;45(4):1313-1320.

7. G.P.K. Marasinghe, I.M. Sander, Brian Bennett, Gopal R Periyannan, Ke-Wu Yang, C.A. Makaroff, and M.W. Crowder, "Structural Studies on a Mitochondrial Glyoxalase II", J. Biol. Chem.(2005); 280 (49) 40668 - 40675.

8. Gopal R. Periyannan, Patrick Shaw, Tara Sigdel and Michael W. Crowder, “In vivo folding of recombinant metallo-b-lactamase L1 requires the presence of Zn(II)”, Protein Science (2004) 13: 2236- 2243.

9. Anne L. Carenbauer, James D. Garrity, Gopal R. Periyannan, Robert B. Yates, and Michael W. Crowder, “Probing substrate binding to Metallo-β-lactamase L1 from Stenotrophomonas  maltophilia by using site-directed mutagenesis”,  BMC Biochem(2002); 3 (1): 4.

10. Crowder, Michael W., Yang, Ke-Wu, Carenbauer, Anne L., Periyannan, Gopal., Seifert, Mary E.,  “The Problem of Solvent Exposable disulfide when Preparing Co (II)-Substituted Metallo-b-lactamase L1 from Stenotrophomonas maltophilia”,(2001) JBIC 6, 91-99.


                                 
International Proteolysis Society
International Protease Network
MEROPS - The Peptidase Database

ZINC Database
International EPR Society
National Biomedical EPR Center
           Go to Top