Professional Documents
Culture Documents
Bel420 Major Sem2 08
Bel420 Major Sem2 08
3. Two proteins have the same molecular weights. At pH 5.5 both have considerable
secondary structure and are at least 75% a helix. At pH 8.5, one of them loses all
structure and is a random coil; when returned to pH 5.5, its structure is restored. Devise a
procedure for separating these proteins. (3)
)f
A procedure has been devised for purifying 1t particular enzyme using gel
. . chromatography. In an effort to increase the amount of material to be handled, you use a
s pie whose concentration of protein is ten times that normally used with this
procedure. The enzyme activity now elutes principally in the void volume. Explain what
happened. .
;J 2'(i) How ma~y ~pm are a~so~iatedwith 1yCi ?~radioa~tive material? lfthe instrume~t
. used for countmg IS 25% efficIent at detectmg dlSlntegratlOnevents, how many cpm w1l1
i yield? What is Cherenkov counting and for which radioactive material can it be
u~? 0r
(ii) What is the role of SDS and polyacryamide gel in SDS-P AGE electrophoresis?
What is the use of Fergueson's plot? (2+ 1)r-
(iii) Are there matrix peaks in the electrospray mass spectra and in MALDI? Explain.
Name two matrices used and their properties. (3)~