a protein sample complex consists of two proteins, a smaller protein, X
Added by Michael H.
Step 1
--- ** Show more…
Show all steps
Your feedback will help us improve your experience
Sri K and 55 other Biology educators are ready to help you.
Ask a new question
Labs
Want to see this concept in action?
Explore this concept interactively to see how it behaves as you change inputs.
Key Concepts
Recommended Videos
A biochemist is attempting to separate a DNA-binding protein (protein X) from other proteins in a solution. Only three other proteins (A, B, and C) are present. The proteins have the following properties: pl (isoelectric point) | Size Mr --- | --- protein A | 7.4 | 82,000 protein B | 3.8 | 21,500 protein C | 7.9 | 23,000 protein X | 7.8 | 22,000 What type of protein separation techniques might she use to separate (a) protein X from protein A? (b) protein X from protein B?
Sri K.
A biochemist is attempting to separate a DNA-binding protein (protein X) from other proteins in a solution. Only three other proteins (A, B, and C) are present. The proteins have the following properties: pI Size (Mr) Bind to GST? protein A 4.1 46,000 yes protein B 3.8 21,500 no protein C 9.8 45,000 no protein X 4.3 45,500 no What type of protein separation techniques should they use to separate: A) protein X from protein A? B) protein X from protein B? C) protein X from protein C?
Sarah C.
Your lab studies two different protein complexes in yeast, Your Favorite Protein Complex I (YFPC1) and Your Favorite Protein Complex II (YFPC2). Following purification of the complexes from cell lysates, you heat them under different conditions and resolve them using SDS-PAGE and Coomassie staining. Your results are shown here. Lane 1: YFPC1 – SDS + mercaptoethanol Lane 2: YFPC2 – SDS + mercaptoethanol Lane 3: YFPC1 – SDS Lane 4: YFPC2 – SDS (a) You have additional evidence suggesting that YFPC2 in its folded, native state has a MW of ~300 kDa. You hypothesize that YFPC2 is composed of some number of single polypeptide chains with identical primary amino acid residue sequence (i.e., monomers). Based on the data here, how many monomers would form one native complex? Show your work. In addition to the Coomassie staining analysis, you perform Western blotting analysis. You transfer the proteins from identical SDS-PAGE gels to PVDF membranes. (In other words, proteins are resolved in the same order and at the same sizes, but instead of Coomassie staining the gel, you transfer them to PVDF.) Draw your results, indicating lanes 1-4 and sizes on the ladder, under the following treatments. (Assume all steps are performed correctly, and that antibodies perform well for this application with idealized results unless otherwise indicated.) (b) The blot is treated with a primary antibody (mouse IgG) that specifically binds the 10 kDa subunit of YFPC1 and a secondary anti-mouse IgG antibody conjugated a fluorescent probe, and imaged with a fluorescent scanner. (c) The blot is treated with a primary antibody (mouse IgG) that binds all protein-protein intermolecular disulfide bridges (strikingly, it does not recognize intramolecular disulfide bridges within a single polypeptide) and a secondary anti-mouse IgG antibody conjugated a fluorescent probe, and imaged with a fluorescent scanner. Bonus (+2 pt) Sketch your results if you do everything correctly EXCEPT you are in a rush and forget to wash away unbound secondary antibodies from your blot before imaging.
Recommended Textbooks
Biology for AP Courses
Objective Biology for NEET
Introduction to General, Organic and Biochemistry
Transcript
18,000,000+
Students on Numerade
Trusted by students at 8,000+ universities
Watch the video solution with this free unlock.
EMAIL
PASSWORD