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OF THE THESIS MOLECULAR CHARACTERIZATION OF GcC8 ALPHA, THE FUNCTIONAL HOMOLOGUE OF HUMAN C8 ALPHA IN THE SHARK, GINGLYMOSTOMA CIRRATUM by Lydia Tatiana Aybar Florida International University, 2010 Miami, Florida Professor Charles H. Bigger, Major Professor The focus of this study is to elucidate the components of the nurse shark (Ginglymostoma cirratum) membrane attack complex (MAC), specifically complement component C8a (GcC8u). Nurse shark C8a gene was cloned, sequenced, and analyzed and Western blot analysis performed to identify components of shark MAC. GcC8a consists of 2341 nucleotides that translate into a 589 amino acid sequence that shares 41.1% and 47.4 % identity with human and xenopus C8a, respectively. GcC8a conserves the MAC modular architecture and cysteine-rich backbone characteristic of complement proteins, including the cysteine residue that forms the C8a-y bond as well as the indel that is unique to C8a. Conservation of MAC protein structure is evident from crossreactivity of antihuman-MAC antibodies with shark serum proteins in Western blots which confirmed the presence of C8 and C9-like proteins in shark serum, however, did not resolve the question of whether C6 and/or C7 like proteins are present in shark.

Molecular characterization of GcC8 alpha, the functional homologue of human C8 alpha in the shark, Ginglymostoma cirratum