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美國布魯克海文儀器公司>技術(shù)文章>測量應(yīng)用案例-20220310

技術(shù)文章

測量應(yīng)用案例-20220310

閱讀:89          發(fā)布時間:2022-3-24
 

文獻名: Structural Analysis of the cl-Par-4 Tumor Suppressor as a Function of Ionic Environment

 

 

作者 Krishna K. Raut,Komala Ponniah andSteven M. Pascal

 Department of Chemistry and Biochemistry, Old Dominion University, Norfolk, VA 23529, USA

 

 

摘要:Prostate apoptosis response-4 (Par-4) is a proapoptotic tumor suppressor protein that has been linked to a large number of cancers. This 38 kilodalton (kDa) protein has been shown to be predominantly intrinsically disordered in vitro. In vivo, Par-4 is cleaved by caspase-3 at Asp-131 to generate the 25 kDa functionally active cleaved Par-4 protein (cl-Par-4) that inhibits NF-κB-mediated cell survival pathways and causes selective apoptosis in tumor cells. Here, we have employed circular dichroism (CD) spectroscopy and dynamic light scattering (DLS) to assess the effects of various monovalent and divalent salts upon the conformation of cl-Par-4 in vitro. We have previously shown that high levels of sodium can induce the cl-Par-4 fragment to form highly compact, highly helical tetramers in vitro. Spectral characteristics suggest that most or at least much of the helical content in these tetramers are non-coiled coils. Here, we have shown that potassium produces a similar effect as was previously reported for sodium and that magnesium salts also produce a similar conformation effect, but at an approximately five times lower ionic concentration. We have also shown that anion identity has far less influence than does cation identity. The degree of helicity induced by each of these salts suggests that the “Selective for Apoptosis in Cancer cells” (SAC) domain—the region of Par-4 that is most indispensable for its apoptotic function—is likely to be helical in cl-Par-4 under the studied high salt conditions. Furthermore, we have shown that under medium-strength ionic conditions, a combination of high molecular weight aggregates and smaller particles form and that the smaller particles are also highly helical, resembling at least in secondary structure, the tetramers found at high salt.

 

關(guān)鍵詞: intrinsically disordered protein (IDP)prostate apoptosis response-4 (Par-4)tumor suppressorcircular dichroism (CD) spectroscopydynamic light scattering (DLS)aggregationcoiled-coilcaspase

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