Proteins - Complete analysis of peptide compositon

8 important questions on Proteins - Complete analysis of peptide compositon

What are the current parameters used to describe a hydrolysate and on what does it not give sufficient information?

Currently used overall parameters

  • Degree of hydrolysis (DH)
  • Extent of degradation of intact proteins
  • Peptide molecular weight distribution


Do not provide sufficient information to
  1.   fully describe the composition of a hydrolysate
  2.   understand the mechanism of hydrolysis

Why is the proteomics method not useful to determine analyse the peptides in food?

To expensive and to much work because there are to many different proteins.
Also there are no specific enzymes present and they are able-free.

How was the peptide annotation of beta-lactoglobuline determent by a pro-tease specific to glutamic acid E and espartic acid D?

It was known that after the enzyme treatment you have 44 peptides. These peptides are analysed by a UV chromatography, MS and a MS2.

This is done for the protein at different points in time in order to follow the hydrolysis.
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How can you analyse the quality of the peptide annotation?

By the amino acid sequence coverage but more by the peptide sequence coverage.
You know what peptides you should find and you know what you did find. This difference will tell you how well the method worked for you annotation.

Can you better use UV or MS for quantification?

UV because it is constant and you can calculate absorption coefficient.

What are the problems with using MS for quantification?

  • You have ionization so you dont know the slope of the peptides and it can only be found with standers which are expensive and cost a lot of time.
  • Variation of angle in which is sprayed an influence the quantification
  • There is a mass charge ratio so this can change from day to day so every day you need to calibrate.
  • Mass can change due to adducts

Why was it sad that you can not use UV for quantification with LC?

Because the uv absorbance is measured in a different form then whit a simple cuvet. It is measured in a cell and for this you need a conversion factor called K(cell) and this is different for every single machine. However it can be calculated so you are able to used the UV in quantification.

How did they they test the consistency of peptide sequence coverage?

They sum the total annotated concentration of each AA and now you can compare this sum with how much of this AA should be present.

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