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SUBRAMANIAM'S STUDENT RESEARCH LAB
Julainna Allport '05
Christopher Butts '04
DIABETIC HYPERGLYCEMIA AND STRUCTURAL MODIFICATIONS OF HUMAN SERUM ALBUMIN
Julianna Allport '05 and Ram SubramaniamMy research this summer focused on how at the biochemical level, diabetic hyperglycemia (elevated blood sugar) leads to complications in diabetic patients, such as retinopathy and cardiovascular disease. Specifically, I worked with the most abundant plasma protein in human serum, human serum albumin (HSA), and observed how when added to HSA in typical human in vivo conditions, five different sugars, (methylglyoxal, glyoxal, glucose, ribose, and glyceraldehyde), modified the structure of HAS. Since structure and function of proteins are closely intertwined, if the structure of the HSA protein is modified when the sugars are added to it, then the function of the protein is also modified. If the function of the HSA is modified, the transport of specific ions and substances in human serum is also altered, and this alteration is what biochemists believe leads to the development of diabetic complications such as those mentioned above.
One way to observe structural modifications of proteins is to look at their hydrophobicity (the degree to which the protein fears water). The hydrophobicity can be deduced by measuring the net charge of the protein. After preparing the normal and sugar-modified HSA samples I used a technique called capillary electrophoresis to measure the net charges of the samples relative to one another. My results for a majority of the samples were excellent and I was able to draw the conclusion that sugars do modify the structure and thus function of human serum albumin by altering the net charge and hydrophobicity of the protein.
Student stipend provided by a grant from Merck/American Association for the Advancement of Science
PROPERTIES OF HUMAN SERUM ALBUMIN
Christopher Butts '04 and Ram SubramaniamThis research project I undertook this summer involved studying various properties of the blood protein Human Serum Albumin (HSA). Properties such as, hydrophobicity, Cu2+ binding ability, and cyanogen bromide segmentation were studied. The protein was also studied after modification by different sugars. The purpose for studying these properties is to eventually find out whether or not HSA or any part of the protein has antioxidant properties that could be used to battle negative side effects of diabetes. If HSA, or a segment of the protein, is determined to be a good antioxidant then it can be used in fighting diabetic side effects such as inflammation.
In my research I used both high performance liquid chromatography (HPLC) and an isothermal titration calorimeter (ITC). The HPLC was used to test the hydrophobicity of HSA and different combinations of sugars with HSA. Cu2+ binding ability was tested by using the ITC to test the same combinations of sugars with HSA as well as different HSA variations. The results for both tests showed some promising information towards the end of research, however further investigation is needed. Also the HSA that was segmented by cyanogens bromide was also collected by different protein segments and stored for further research.
Stipend support for Christopher Butts was provided by the Ralph E. Hansmann Science Students Support Fund