The oligomerization of Influenza A Virus matrix 1 (M1) protein was analyzed usin

Question

The oligomerization of Influenza A Virus matrix 1 (M1) protein was analyzed using affinity chromatography followed by gel filtration. (A) Purified recombinant M1 (with a C-terminal His6-tag) from nickel affinity chromatography eluted in peak 4. The fraction containing peak 4 (from panel A) was collected. SDS-PAGE analysis determined that the peak 4 fraction was 100% pure. (B) Half of the collected fraction was applied to a Superdex 200 HR 10/30 gel filtration column at pH 7.4 (physiological pH). (C) Half of the collected fraction was applied to a Superdex 200 HR 10/30 gel filtration column at pH 5.0 Which peak in panel B contains protein(s) with the highest molecular weight? What experimental condition causes multiple peaks in panel B to elute as a single peak in panel C? Propose a reasonable scientific hypothesis about the biochemical basis for multiple peaks in panel B, but a single peak in panel C.

Explanation / Answer

In panel B peak D contains proteins with the highest molecular weight.

The change in the pH of the buffer causes multiple peaks in panel B to elute as a single peak in panel C.

The protein is stable in the pH of 5.0 rather than 7.5, The protein looses its quaternary structure and forms aggregates which has made it to elute as different molecular weight peaks in panel B.

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