Chapter 5: Q15P (page 127)
A pentapeptide has the sequence
Short Answer
The mass of this peptide, as determined by mass spectrometry, to three significant figures is
Chapter 5: Q15P (page 127)
A pentapeptide has the sequence
The mass of this peptide, as determined by mass spectrometry, to three significant figures is
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Get started for free(a) The ESI-MS spectrum below was obtained for hen egg-white lysozyme (HEWL). Using peaks 5 and 6, calculate the molecular mass of HEWL (see Sample Calculation 5-1).
[http://www.astbury.leeds.ac.uk/facil/MStut/mstutorial.htm. Published by A.E. Ashcroft, Astbury Centre for Structural Molecular Biology, University of Leeds (2000)]
(b) What is the charge on the ion that makes peak 5?
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?
You must cleave the following peptide into smaller fragments. Which of the proteases listed in Table 5-4 would be likely to yield the most fragments? The fewest?
NMTQGRCKPVNTFVHEPLVDVQNVCFKE
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?
You are trying to purify a protein that is soluble in a solution of 2 M ammonium sulfate. After centrifugation to remove other proteins that have precipitated at this high salt concentration, you recover the supernatant to assay the target proteinโs activity in a cell culture system.
(a) Explain why the cells die when incubated with the supernatant.
(b) What procedure could you perform to correct the problem? (Hint: See Section 2-1D).
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