- 2012-05 (x)
- Vander Schrier, Ann (x)
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Study of urban community gardens in the City of Cleveland, Ohio.
Show moreBetaine homocysteine methyltransferase-2 (BHMT-2) is a zinc metalloenzyme that plays an important role in the regulation of the homocysteine metabolism. It uses S-methylmethionine (SMM) as a methyl donor for the methylation of homocysteine. BHMT-2 is a 73% homologous to the protein BHMT, which catalyzes the remethylation of betaine and homocysteine to form dimethylglycine and methionine. BHMT is abundant in the human liver, kidney, and lens of the eye. BHMT (and likely BHMT-2) are involved in regulating homocysteine and S-adenosyl-L-methionine (SAM, AdoMet) levels. The goal of this study was to generate recombinant BHMT-2 for structural and functional studies. Escherichia coli BL21 (DE3), were transformed with the expression construct coded for BHMT2, fused with a N- terminal His-tag, and then were grown. Recombinant BHTM-2 protein was purified using 6xHis-tag affinity chromatography. Also, size exclusion chromatography was used to purify His-BHMT-2. The dimer form of His-BHMT-2 protein was obtained as a result in the study. By the analyzed results of this study, the protein BHMT-2 can be purified by different methods of chromatography. BHMT-2 polymorphisms may contribute to BHMT-2 enzyme variants with different levels of activity, thus leading to different levels of SAM (aka AdoMet), which influences acetaminophen sensitivity/response and thus liver toxicity. However, further experiments are needed to make structural studies (crystallography) and functional (enzyme assays) to compare the WT and variant enzymes (corresponding to genetic polymorphisms in the gene). It is very important to make further studies in way to determine the mechanism and function of this protein in humans.
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Show moreActivating mutations in SHP-2 tyrosine phosphatase implicated in cell signaling processes have been identified in various childhood leukemias and sporadic solid tumors. However, the molecular mechanisms by which the activating mutations in SHP-2 induce malignancies are not fully understood. In this study, we investigated potential involvement of SHP-2 in chromosome segregation during mitosis, dysregulation of which leads to genome instability and malignancies. We determined localization of SHP-2 in the kinetochore, a crucial protein complex structure connecting microtubules and chromosomes. We found that SHP-2 localized at the kinetochore. Our findings suggest that SHP-2 may be involved in chromosome segregation and can help to answer how SHP-2 mutations can induce genetic instability and thereby malignancies.
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