Department of biochemistry and Molecular biology
In recognition of the critical role that it plays in biomedical research across many disciplines, MUSC has made a significant investment in structural biology and the Department of Biochemistry houses most of its major facilities. The X-ray crystallography is located on the 5th Floor of the Basic Sciences building and contains Riguku diffraction equipment, a crystallization lab with dispensing robotics and an imaging system, and a molecular graphics facility. The X-ray facility also provides remote access to the SER-CAT synchrotron beamlines at the Advanced Photon Source, near Chicago. The NMR facility has undergone significant expansion recently and now offers 600 and 850 MHz magnets for biomolecular studies, in addition to a 400 MHz instrument used for small-molecule work. The Department also houses the Protein Production Laboratory (currently on the 7th floor of BSB, but soon moving to the 5th floor), established to support structural biology. This lab offers scale-up protein production and biophysical characterization (circular dichroism and dynamic light scattering).
Lipidomics Shared Resource
Lipidomics Shared Resource (LSR) at MUSC represents the only scientific shared resource of its kind; providing service to the Medical University of South Carolina, medical institutions throughout the world in addition to pharmaceutical companies. LSR is composed of synthetic and analytical units providing expertise, mentoring, synthetic molecular tools and analytical methodology to study significance of bioactive sphingolipids.
The Lipidomics Shared Resource builds on unique expertise at MUSC in sphingolipid biology, chemistry and analysis and their role in signal transduction and cell regulation. This expertise supports the facility's pivotal role in MUSC's COBRE in Lipidomics and Pathobiology.
Sphingolipid metabolism assumes a key role in the complex mechanisms regulating cellular stress responses to environmental changes. Sphingolipids are recognized as a key components regulating fundamental cell biology processes acting as modulators for transmembrane signaling and mediators for cellular interaction. Diversity of bioactive sphingolipids and their interconnected metabolism provide a network of pathways regulating intra- and inter-cellular signaling and function. Key sphingolipid metabolites: ceramides (Cer), sphingosine (Sph) and sphingosine 1-phosphate (S1P), act as bioactive molecules that mediate and/or regulate various cell responses including cell growth, cell death, inflammation, cell migration, and cell senescence. Dysfunctions in the metabolism of these sphingolipids contribute to pathology of diseases. To understand how sphingolipids biosynthesis and turnover regulate cell behavior, it is essential to determine the levels of these bioactive molecules in order to examine their effects on cell function.
This emphasizes the needs to analyze sphingolipid components, examine sphingolipid chemistry and regulation of sphingolipid metabolic pathways. Monitoring changes in sphingolipid composition under normal and disease environments may provide one of the missing links in the search for a novel and effective therapy.
Facilities, Equipment, Services
The Nucleic Acid Analysis Facility of the Biochemistry Department provides automated DNA sequence analysis to members of the MUSC community. The service includes performing thermal cycling sequencing reactions on investigator-provided DNA templates followed by polyacrylamide gel electrophoresis, laser-induced fluorescence detection analysis of the gel, data collection and analysis. Both plasmid DNA and PCR fragments can be sequenced and a variety of primers are provided by the facility. Custom, investigator supplied primers can also be used for sequence analysis. Sequence data are provided to investigators as four-color electrophoregram plots and as text files delivered via E-mail or on a computer disc.
The Facility also educates researchers regarding the capabilities of DNA analysis technologies and assists in developing and applying these technologies for specific projects.
At present, the Nucleic Acid Analysis Facility is located in room # 217, sharing space with the GCRC molecular lab.
Currently the Facility uses an ABI Prism 377 Genetic Analyzer to collect and analyze the sequencing data as well as an ABI 310 Genetic Analyzer. The ABI 310 analyzer can also be used for- microsatellite analysis and DNA fragment sizing for RFLP/AFLP analysis. GeneScan software is available for analysis of fragment sizes. Recently, the facility resources expanded due to acquisition of an additional GE Healthcare MegaBase 500 instrument, which more than doubles the production capacity.