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​A purified 150 kDa protein species from a gel filtration column was run on a 2-dimensional SDS-PAGE as shown below:​​What is the likely form of the 1
Question

A purified 150 kDa protein species 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 protein 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 that has a disulfide bond.

Correct option is B

Sol.

  • The gel filtration shows a 150 kDa species, indicating a complex or multimer.

  • In SDS-PAGE without DTT, the complex remains intact, showing one or few bands corresponding to subunits linked by covalent bonds (disulfide bridges).

  • Upon SDS-PAGE with DTT, disulfide bonds are reduced, and the protein complex dissociates into smaller subunits, visible as separate bands.

  • This implies covalent linkage (disulfide bonds) between subunits, confirming multiple proteins linked covalently.

Information Booster:

  • DTT (dithiothreitol) reduces disulfide bonds, disrupting covalent linkages.

  • Non-covalent interactions would be disrupted by SDS alone, so complexes linked non-covalently wouldn't maintain structure in SDS-PAGE without DTT.

  • 2D SDS-PAGE with and without reducing agents helps identify protein complexes and subunit structure.

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