GVE

I created pH dependence on log(Vmax/Km) graph, but I'm not sure how I can get pKa values from it, because it's form is strange. Then I created pH dependence on log(Vmax) graph: and painted lines to find pKa values. Am I doing it right?

I have information Km and Vmax dependence on pH: pH 6.0 6.3 6.5 7.0 7.5 8.0 KM (mM) 0.120 0.163 0.216 0.250 0.210 0.160 Vmax 0.430 0.480 0.526 0.530 0.450 0.313 and I have to determine which amino acids are in the active site. Which is a better way to find pKa and determine the amino acids? To look at logKm dependence on pH, or at logVmax dependence on pH? Thank you.

So, if 50 microlitres shows concentration of 1,7mg/ml, then it means that 2mL would be 1,7mg/ml*2ml=3,4mg? Am I right?

How to count the amount of protein in 2ml of extract, when 50microlitres of this extract were diluted in 3ml of buffer and concentration of protein was 1,7mg/ml. Thank you.

Aspartate aminotransferase (AAT) enzyme was being purified from pigs heart. Tissues were homogenated and cells were eliminated with gel filtration. Nucleic acids were precipitated with polyethylenimine and extract1 was received with volume 2dm3. Then 50microLitres of this extract1 were added to 3cm3 buffer and optical density was measured at 280nm (1cm light path) and the value of it was 1,7. How should I count the amount of protein in extract1?