This project focuses on the roles of human hyaluronidase-1 (hHyal-1) andhyaluronidase-2 (hHyal-2) in the mediation of the biological functions assumedby the extracellular matrix polysaccharide hyaluronan (HA). HA serves as the'glue' that binds cells together. In order for cells to move in or out of the matrix,HA must be fragmented by hHyals. Repair of injured tissue requires cellmovement within the matrix, as does the escape of an abnormal cell(metastasis). HA fragments are detected by neighboring cells, and anintracellular response occurs via signal transduction pathways. The signalreceived is dependent on the size of the HA fragments as produced by the Hyals.Of particular medical importance, hHyal-1 and hHyal-2 play key roles in tissueinflammation and in cancer, and therefore are excellent targets for thedevelopment of novel anti-cancer and anti-inflammatory therapeutics. Untilrecently, structure-function studies of these enzymes have not been possibleowing to the technical challenge associated with high yield production of theactive N-glycosylated hHyals. A method for hHyal-1 and hHyal-2 preparation hasbeen recently developed in the Herzberg lab and the crystal structure of hHyal-1has been determined. The stage is now set to employ in vitro methods toaccurately define hHyal-1 and hHyal-2 catalytic function and inhibition, and theirinteractions with protein partners, CD44 - the hyaluronan biding protein, RONreceptor tyrosine kinase that is regulated by hHyal-2, and Jaagsiekte sheepretrovirus envelope protein that activates RON and uses hHyal-2 to attach to thehost cell. The research plan set forth in this proposal will illuminate at the atomiclevel the molecular mechanisms underlying the complex cellular processescontrolled by the hyaluronidases and will provide the structural foundation for thedevelopment of new therapeutics.

Public Health Relevance

Hyaluronidase-1 and hyaluronidase-2 are crucial to the integrity and functioning of the extracellular matrix through their enzymatic activity that controls the turnover of the extracellular polysaccharide hyaluronan (HA). The HA turnover is delicately balanced and heightened hyaluronidase activity leads to inflammatory diseases and cancer progression and invasion. This project seeks to characterize the human hyaluronidases and their interactions with cellular partner proteins in vitro to shed light on their in vivo function and to lay the foundation for development of the hyaluronidases as anti cancer and anti inflammatory drug targets.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Special Emphasis Panel (ZRG1-IDM-Q (03))
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Flicker, Paula F
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University of Maryland College Park
Other Domestic Higher Education
College Park
United States
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Chao, Kinlin L; Gorlatova, Natalia V; Eisenstein, Edward et al. (2014) Structural basis for the binding specificity of human Recepteur d'Origine Nantais (RON) receptor tyrosine kinase to macrophage-stimulating protein. J Biol Chem 289:29948-60
Chen, Chen; Gorlatova, Natalia; Herzberg, Osnat (2012) Pliable DNA conformation of response elements bound to transcription factor p63. J Biol Chem 287:7477-86
Chao, Kinlin L; Tsai, I-Wei; Chen, Chen et al. (2012) Crystal structure of the Sema-PSI extracellular domain of human RON receptor tyrosine kinase. PLoS One 7:e41912
Gorlatova, Natalia; Chao, Kinlin; Pal, Lipika R et al. (2011) Protein characterization of a candidate mechanism SNP for Crohn's disease: the macrophage stimulating protein R689C substitution. PLoS One 6:e27269
Chen, Chen; Gorlatova, Natalia; Kelman, Zvi et al. (2011) Structures of p63 DNA binding domain in complexes with half-site and with spacer-containing full response elements. Proc Natl Acad Sci U S A 108:6456-61