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NMR Structure of Mistic, a Membrane-Integrating Protein for Membrane Protein Expression

Structure Determination of Membrane Proteins. Structural determination of soluble proteins has minimal restraintsStructural determination of Membrane Proteins, however, has a couple of restraints: 1. Production of high enough yield of protein2. Crystallization. Characteristics of an ideal fu

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NMR Structure of Mistic, a Membrane-Integrating Protein for Membrane Protein Expression

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    1. NMR Structure of Mistic, a Membrane-Integrating Protein for Membrane Protein Expression By: Niloufar Safvati

    2. Structure Determination of Membrane Proteins Structural determination of soluble proteins has minimal restraints Structural determination of Membrane Proteins, however, has a couple of restraints: 1. Production of high enough yield of protein 2. Crystallization

    3. Characteristics of an ideal fusion partner that is specialized in producing recombinant IM proteins An ideal fusion partner should: autonomously traffic its cargo to the membrane, bypassing the translocon and associated toxicity issues retain the characteristics of other successful fusion partner proteins, including relatively small size, in vivo folding, and high stability.

    4. NMR Spectroscopy Can be used as an alternative method to crystallization NMR structure determination of IM proteins has been established only for very small, structurally simplistic IM proteins and for outer membrane bacterial porins New techniques for determining the characteristics of alpha helical IM proteins are therefore necessary

    5. What is Mistic? Mistic is a Bacillus subtilis integral membrane protein that folds into the membrane without the help of a translocon Mistic stands for Membrane-Integrating Sequence for Translation of Integral Membrane protein Constructs It consists of 110-amino acids (13kD)

    6. Why study Mistic? When recombinantly expressed in E. coli, Mistic associates tightly with the bacterial membrane. Surprisingly, Mistic is highly hydrophilic Mistic has most of the characterizations for being an ideal partner in the production of high-yields of integral membrane proteins

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