Cific cAMPbinding proteins, is somewhat higher in comparison towards the other folks. Our data points out that timolol treatment includes a useful action of antioxidant defense mechanism enzymes around the pancreas of STZinduced diabetic rats. POS430 Enhancing RNA Modification Mapping Sequence Coverage by means of a Nonspecific RNase U2-E49A VariantBeulah Solivio1, Balasubrahmanyam Addepalli1, Patrick Limbach1 University of Cincinnati, USAThe place of RNA post-transcriptional modifications gives a clue on the possible function of a certain modification. The approach employed to ascertain the sequence place of a modification is known as post-transcriptional modification mapping. This strategy makes use of ribonucleases that cleave lengthy RNA sequences into fragments that may be detected by the mass spectrometer. This process requires the usage of multiple enzymes which have unique cleavage specificities to characterize as a great deal sequence coverage as you can. At present, RNase T1 could be the only base-specific enzyme accessible in the market place. The goal of this study would be to discover the potential of RNase U2, a purine distinct enzyme with a preference to cleave adenosine in the 30 -end, as a tool for RNA modification mapping. Here we demonstrate the useABSTRACTSof RNase U2-E49A, a nonspecific variant of RNase U2 that generates random lengthy sequences, enabling RNA modification mapping. We are going to discuss the positive aspects and setbacks of using this enzymes. Our existing work also explores enhancing the adenosine specificity of RNase U2 by phage display mutagenesis. POS433 Phosphate Affects The Quaternary Structure of Alanine Racemase from Mycobacterium TuberculosisJohn C Ford1, Shannon A. Stirling1, Jaeju Ko1, Sudipta Majumdar1 Indiana University of Pennsylvania, USAIn Mycobacterium tuberculosis (MT), the causative agent of tuberculosis, active alanine racemase (ALR) exists as a dimer, with two active websites, each and every formed around the interface between the individual monomers. A total understanding with the interaction amongst the two monomers could offer insight into approaches to inactivate the dimeric ALR. We investigated the degree of association of ALR in by indicates of higher functionality size exclusion liquid chromatography (HPSEC), In buffers containing phosphate over the pH selection of 52, MT ALR eluted with a retention volume constant using a molar mass of about 70 kDa, i.e., as a monomer-dimer equilibrium mixture. On the other hand, in pH eight.0 borate buffer, MT ALR eluted having a retention volume constant using a molar mass of 80 kDa, i.e., the expected dimer. Additional, when phosphate was added for the borate buffer, the HPSEC retention volume of MT ALR decreased. These results are constant with MT ALR possessing a phosphate binding-site that inhibits the dimerization vital for its activity. POS441 Mechanisms of activation and substrate recognition by PINK1, a ubiquitin kinase implicated in mitochondrial excellent control and Parkinson’s diseaseJean-Franc Trempe1 is 1 McGill University, CanadaMutations in Parkin and PINK1 lead to an early-onset autosomal recessive form of Parkinson’s disease (PD). PINK1 is usually a kinase that acts as a sensor of mitochondrial harm and initiates Parkin-mediated quality handle. Recruitment and activation of Parkin by PINK1 results in the ubiquitination of outer mitochondrial membrane proteins and subsequent autophagic clearance of the damaged organelle. PINK1 then phosphorylates ubiquitin at Ser65 around the mitochondrial outer membrane (Tang et al.