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Chapter 5: Q14CP (page 119)
What types of information can be retrieved from a protein sequence database?
The protein sequence database retrieve protein sequence records by gene name, organism, author name, accession number and a variety of other text terms.
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Draw a different phylogenetic tree based on the data in Problem 19.
Explain how the number of amino acid differences between homologous proteins can be used to construct a phylogenetic tree.
Question: Purification tables are often used to keep track of the yield and purification of a protein. The specific activity is a ratio of the amount of the protein of interest, in this case Mb, obtained at a given step (μmol or enzyme units) divided by the amount (mg) of total protein. The yield is the ratio of the amount of the protein of interest obtained at a given step (μmol or enzyme units) divided by the original amount present in the crude extract, often converted to percent yield by multiplying by 100. The fold purification is the ratio of the specific activity of the purified protein to that of the crude preparation.
(a) For the purification table below, calculate the specific activity, % yield, and fold purification for the empty cells.
(b) Which step—DEAE or affinity chromatography—causes the greatest loss of Mb?
(c) Which step causes the greater purification of Mb?
(d) If you wanted to use only one purification step, which technique would you choose?
Electrospray ionization mass spectrometry (ESI-MS) of proteins involves creating positively charged ions of the protein and separating them according to their mass-to-charge ratio (m/z).
(a) What causes the different positive charges on different particles of the protein?
(b) The amino acid composition (in numbers of residues per chain) of hen egg-white lysozyme (HEWL) is as follows:
P 2 Y 3 N 14 H 1
D 7 M 2 L 8 E 2
C 8 R 11 G 12 F 3
A 12 I 6 K 6 V 6
S 10 W 6 T 7 Q 3
What is the maximum positive charge that can be present on a HEWL ion?
Explain how you could use a column containing carboxymethyl groups to separate serum albumin and ribonuclease A(see Table 5-3).
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