C 1 -tetrahydrofolate (THF) synthase is a trifunctional enzyme found in eukaryotes that contains the activities 10-formyl-THF synthetase, 5,10-methenyl-THF cyclohydrolase, and 5,10-methylene-THF dehydrogenase. The cytoplasmic isozyme of C 1 -THF synthase is well characterized in a number of mammals, including humans; but a mitochondrial isozyme has been previously identified only in the yeast Saccharomyces.

Mammalian mitochondrial C 1 -tetrahydrofolate (THF) synthase (MTHFD1L gene product) is a monofunctional 10-formyl-THF synthetase, lacking the 5,10-methylene-THF dehydrogenase and 5,10-methenyl-THF cyclohydrolase activities typically found in the trifunctional cytoplasmic proteins. Here we report the submitochondrial localization of epitope-tagged human mitochondrial C 1 -THF synthase expressed in Chinese hamster ovary cells. Mitochondrial fractionation experiments show that human mitochondrial C 1 -THF synthase behaves as a peripheral membrane protein, tightly associated with the matrix side of the mitochondrial inner membrane. Inner mitochondrial membrane association was also observed for the endogenous mitochondrial C 1 -THF synthase in adult rat spleen. We also purified and characterized the recombinant protein product (short isoform) of the alternatively spliced short transcript of the mitochondrial isozyme. Methylene-THF dehydrogenase assays confirmed that the short isoform is not enzymatically active. The purified short isoform was used in the production of polyclonal antibodies specific for the mitochondrial isozyme. These antibodies detected endogenous full-length mitochondrial C 1 -THF synthase in mitochondria from adult rat spleen and human placenta, confirming the expression of the mitochondrial isozyme in adult mammalian tissues.

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Floating columns are columns which rest on beams and do not have foundation, are common multi storey buildings which are proposed to accommodate parking at ground floor or open halls at higher floors. These columns have discontinuities in the load transfer path and are designed for gravity loads but these buildings are not designed for earthquake loads. So these buildings are unsafe in seismic prone areas. In the study, the effect of varying the location of floating columns floor wise and within the floor of multi storied RC building on various structural response quantities of the building using response spectrum analysis is studied in the software ETABS 2015. The main objective here is to study the seismic response of building with floating columns and to find out the most suitable configuration for providing floating columns. Various parameters such as total base shear force, storey displacement, storey drift, story acceleration of a building are studied with respect to different configurations of floating columns. Also, the critical configuration is taken and made it resistant against lateral loads by providing shear wall