2. Protein pl vs. pH. Estimate the approximate net charge (as either t indicated

Question

2. Protein pl vs. pH. Estimate the approximate net charge (as either t indicated pH values or 0) on the following proteins at the Protein pl Approximate Oarge at pH 4.00 5.00 6.00 7.00 8.00 9.00 10.00 5.43 6.82 7.50 8.64 9.25 3. Protein Purification Tables. Complete the following purification table for a hypothetical enzyme fold purification purification step fraction total fraction total volumeprotein [protein] activity (mL) specific yield activity (mg) (mg/mL) units (mU) (mU/mg) clarified cell extract 225 ion-exchange affinity (Ni-IMAC) 50 12 2,277 689 95 48,529 35,278 20,345 1 Show your calculation for % yield HERE 2 Show your calculation for fold purification after Ni-IMAC HERE 4. Chromatography Resins. Complete the molecule structure at each nitrogen atom, so that the key group on each of the named chromatography resins is accurately represented. Finally, label each as either an "anion-exchanger" or "cation-exchanger" Q-Sepharose DEAE-Cellulose agarose bead cellulose bead

Explanation / Answer

A

Remember following points for these questions

1. When ph >pI , charge on protein is positive .

2. When pH<pI ,charge on protein is negative .

3. When pH=pI , charge on protein is zero

Protein pI ph 4 5 6 7 8 9 10

A

5.43 + + - - - _ _ B 6.82 + + + - _ _ _ C 7.50 + + + + _ _ _ D 8.64 + + + + + _ _ E 9.25 + + + + + + _

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