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    ​A purified 150 kDa species obtained from a gel filtration column was run on a 2-dimensional SDS-PAGE as shown below:​​What is the likely form of the
    Question

    A purified 150 kDa species obtained from a gel filtration column was run on a 2-dimensional SDS-PAGE as shown below:

    What is the likely form of the 150 kDa species from this observation?

    A.

    There are at least two proteins that are linked through non-covalent interactions.

    B.

    There are at least two proteins in the complex that are linked through covalent bonds.

    C.

    There are two proteins in the mixture without forming a complex.

    D.

    There is only one protein and it has a disulfide bond.

    Correct option is A

    The correct option is (a)

    EXPLANATION

    Proteins linked by non-covalent interactions can sometimes form stable complexes that resist dissociation during the first SDS-PAGE, especially if the conditions are mild or partial.
    DTT can help break certain interactions or facilitate complete denaturation in the second dimension.
    Hence, the observation fits a complex stabilized by non-covalent interactions that remains intact during the first SDS-PAGE but dissociates into components in the second SDS-PAGE with reducing conditions.

    Incorrect options-

    Option (b) : Covalent bonds here mainly refer to disulfide bonds between protein subunits. If proteins were linked by disulfide bonds, SDS alone wouldn’t break them, so the complex would remain as one band in both dimensions until DTT was added. But the separation seen here indicates more complex dynamics.
    Option (c) : They would separate into two bands even in the first dimension.
    Option (d) : A single protein with disulfide bonds wouldn’t separate into two distinct spots in the second dimension.


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