4. What can you conclude about each of the proteins in this mixture based on the

Question

4. What can you conclude about each of the proteins in this mixture based on the molecular weight measurements provided? Give your responses in the table below: Protein Molecular name weight (in kDa) kDa) as determined protein than one subunit, Molecular weight (in Does this If there is more as determined by SDS PAGE with consist of what is the most by Native gel DTT treatment electrophoresis more than likely subunit one peptide composition for subunit?this protein in (yes or no) the native conformation? Abelp 180 60 & 30 Defip 65 65 Ghilp 60 20 Jkl1p 100 100 Mnolp 150 25 Pqrlp 200 100

Explanation / Answer

Native PAGE helps in determination of molecular weight of the complete protein. However during SDS PAGE with DTT ( reducing agent), the oligomeric proteins get separated into their subunits. So, if the native PAGE and SDS- PAGE of a given protein shows a single band of same size, it means that the protein comprises of a single sub unit. On the other hand if the SDS -PAGE shows variation in the banding pattern (i.e. different size and number of bands as compared to native PAGE), it indicates the protein contains 2 or more sub units.

1. Abc 1p - native 180 kd band, SDS - PAGE, 60 kd, 30 kd bands.

So, yes, it consists of more than 1 subunits.

It may either contain 4 subunits - 2 subunits of 60 kd, 2 subunits of 30 kd or it may contain 5 subunits - 1 subunit of 60 kd and 4 subunits of 30 kd each.

2.Def1p - native and SDS PAGE - 65 kd

No, it does not contain more than 1 subunits.

3. Ghi1p - native -60, SDS PAGE 20

Yes, it contains more than 1 subunits.

It consists of 3 subunits of 20 kd each.

4. Jk1p- native and SDS PAGE - 100 kd

No, it does not have more than 1 subunits.

5. Mno 1p - native - 150 kd, SDS PAGE - 25 kd

Yes, it has more than 1 subunits.

It consists of 6 subunits of 25 kd each.

6. Pqr1 p - native 200, SDS PAGE -100

Yes, it contains more than 1 Subunits.

It consists of 2 subunits of 100 kd each.

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