The following gel are Rubisco bands from Crude Homogenate (CH), Low speed supern
ID: 200477 • Letter: T
Question
The following gel are Rubisco bands from Crude Homogenate (CH), Low speed supernatant (LSS), High-speed supernatant (HSS), and High-speed pellet (HSP). Rubisco consist of 16 subunits (=550KDA), made up 8 large subunits (52kd) and 8 small subunits (13kd). Using the gel results determine the MW from relative mobility of each well and In a paragraph answer the following question
Which fractions do you expect to contain the most rubisco protein and why?
Do you anticipate a similar distribution of small and large subunits across the fractions tested and why?
Is there any other information that can be obtained?
MW: 10 ul, R: 20ul, CH,LSS,HSS,& HSP was 40ul. ONLY DO 1-6 wells (labeled ones).
R CH LSS HSS CH LSS HSS HSP 250 150 100 75 250 150 100 75 50 50 37 37 25 25 20 20 15 15 10 10Explanation / Answer
Ans-
Most Rubisco protein is coming in the HSS - high speed supernatant fraction
Because as it can be observed from gel that most prominent band of larger subunit i.e 52 kD and smaller sub unit 13 kD coming well, where as the intensity of band is not clear in pellet fraction.
And in addition there is an extra band coming around 25 kD which could be denaturation or degradation of larger subunits of Rubisco.
Yes- Similar band pattern is being observed in CH-, LSS also.
The information here obtained is in pellete fraction i.e HSP the extra band which is around 25 kD could be because of degradation or denaturation of Rubosco protein and
also very less intensity of smaller sub unit bands i.e of 13 kds.
some denatured bands piled together at the edge of the gel which could be the breakdown of 13 kd subuntis
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