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Chapter 5: Q5-8P (page 127)

Explain how you could use a column containing carboxymethyl groups to separate serum albumin and ribonuclease A(see Table 5-3).

Short Answer

Expert verified

Albumin has a pI of 4.9 while ribonuclease A has a pI of 9.4. While serum albumin runs through the column, carboxymethyl groups will react with ribonuclease A.

Step by step solution

01

Definition of isoelectric point

The pH of a solution at which the net charge of a protein becomes zero is known as the isoelectric point.

02

Explain the separation of serum albumin and ribonuclease A

Ribonuclease A has a pI of 9.4 and albumin has a pI of 4.9. Due to ribonuclease A having high pl value the Carboxymethyl groups will react with ribonuclease A while serum albumin flows through the column at the same time.

This helps in a column containing carboxymethyl groups to separate serum albumin and ribonuclease A.

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Most popular questions from this chapter

You are using ammonium sulphate to purify protein Q (pI = 5.0) by salting out from a solution at pH 7.0. How should you adjust the pH of the mixture to maximize the amount of protein Q that precipitates?

Separate cleavage reactions of a polypeptide by CNBr and chymotrypsin yield fragments with the following amino acid sequences. What is the sequence of the intact polypeptide?

CNBr treatment

1. Arg鈥揂la鈥揟yr鈥揋ly鈥揂sn

2. Leu鈥揚he鈥揗et

3. Asp鈥揗et

Chymotrypsin

4. Met鈥揂rg鈥揂la鈥揟yr

5. Asp鈥揗et鈥揕eu鈥揚he

6. Gly鈥揂sn

What types of information can be retrieved from a protein sequence database?

You wish to sequence the light chain of a protease inhibitor from the Brassica nigra plant. Cleavage of the light chain by trypsin and chymotrypsin yields the following fragments. What is the sequence of the light chain?

Chymotrypsin

1. Leu鈥揌is鈥揕ys鈥揋ln鈥揂la鈥揂sn鈥揋ln鈥揝er鈥揋ly鈥揋ly鈥揋ly鈥揚ro鈥揝er

2. Gln鈥揋ln鈥揂la鈥揋ln鈥揌is鈥揕eu鈥揂rg鈥揂la鈥揅ys鈥揋ln鈥揋ln鈥揟rp

3. Arg鈥揑le鈥揚ro鈥揕ys鈥揅ys鈥揂rg鈥揕ys鈥揚he

Trypsin

4. Arg

5. Ala鈥揅ys鈥揋ln鈥揋ln鈥揟rp鈥揕eu鈥揌is鈥揕ys

6. Cys鈥揂rg

7. Gln鈥揂la鈥揂sn鈥揋ln鈥揝er鈥揋ly鈥揋ly鈥揋ly鈥揚ro鈥揝er

8. Phe鈥揋ln鈥揋ln鈥揂la鈥揋ln鈥揌is鈥揕eu鈥揂rg

9. Ile鈥揚ro鈥揕ys

10. Lys

You wish to determine the sequence of a polypeptide that has the following amino acid composition.

1 Ala 4 Arg 2 Asn 3 Asp 4 Cys 3 Gly 1 Gln 4 Glu 1 His 1 Lys 1 Met 1 Phe 2 Pro 4 Ser 2 Tyr 1 Trp

(a) What is the maximum number of peptides you can expect if you cleave the polypeptide with cyanogen bromide?

(b) What is the maximum number of peptides you can expect if you cleave the polypeptide with chymotrypsin?

(c) Analysis of the intact polypeptide reveals that there are no free sulfhydryl groups. How many disulfide bonds are likely to be present?

(d) How many different arrangements of disulfide bonds are possible?
See all solutions

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