In clinical studies, Inulin and para-amino hippuric acid given intraveneously until steady state is reached in a 70 kg man. Once steady state was reached, the following measurements were collected: arterial plasma concentration of inulin 0.18 mg/ml, of paraamino hippuric acid 0.21 mg/ml; renal venous plasma concentration of para-amino hippuric acid 0.0014 mg/ml; urine flow 2.2 ml/min; urinary concentration of inulin 11.0 mg/ml, of para-amino hippuric acid 5.5 mg/ml. Calculate the renal venous concentration of inulin.

Total Body Water = 42L

Extracellular Fluid = 14L

Osmolarity = 300mOsmoles/L

Intracellular Fluid = 28L

Enzyme concentrations are often characterized by 'units' of activity per volume.

One enzyme we will be using, Sbfl, is supplied in a concentration of 10,000 units/mL

a. How many are there in 1 uL?

b. How much volume would you need for 20 units?

c. We have 500 units total, how much volume is that?

Enzyme catalysis catalyzes the breakdown of H2O2. Data are:

If the concentration of the enzyme is 4.0x10- 9 mol / L, plot the data from the data and calculate the Michaelis constant and turnover number to find the maximum starting speed?

What kind of enzyme is chymotrypsin? What are the specific substrates of chymotrypsin?

• What features of chymotrypsin make it specific to particular substrates ?

• Can you describe the main features of the reaction mechanism?

• What is a catalytic triad and what is its role in the active site of chymotrypsin?

• What is an acyl-enzyme intermediate?

• What is conformational selection vs. “induced fit”?

Pyruvate Dehydrogenase is a huge multi-enzyme complex in which substrates are channeled from one enzyme to the next. In fact, recent evidence suggests that many citric acid cycle enzymes may interact pass substrates along the pathway and that the components of the electron transport chain associate into an electron transfer super complex. What are the advantages of such an arrangement?

For hemoglobin and myoglobin, discuss the mechanism, similarities, differences, and effects of O₂ binding on the enzymes. What happens to the enzyme structure, and what do these structural changes then cause in terms of functional changes, if any? What role does BPG play for the enzyme and why would this be the case? What about H⁺? How does a baby breathe in the womb?  

**Biochemistry**

Clearly explain how the structure of hemoglobin is intricately connected to its function, using the principle listed below as the outline.

• NCFs rule everything from the structure of biomolecules, to protein ligand binding to enzyme catalysis. (consider protein folding, binding dissociation constants, enzyme mechanisms where binding energy reduces activation energy, etc.)

What is a gene and how does the enzyme that catalyzes transcription find the start of the gene?

What is the DNA chain/thread read in the transcript?

Do the eukaryotes also have only one enzyme that catalyzes the transcription of RNA?

How are the eukaryotic primary mRNA transcripts processed before they can be transported from the cell nucleus to the ribosomes in the cytosol?

Describe the clinical consequences of a genetic or nutritional deficiency in the enzyme or cofactor listed below. Describe the cause of this disease (lack of type of food or genetic inheritance pattern) and its prevalence. Include any theory that describes the molecular mechanisms that connect the lack of enzyme activity to the disease symptoms.

a) hexokinase
b) phosphofructokinase
c) glyceraldehyde 3-phosphate kinase

What are the two general strategies for maintaining the very high degree of fidelity of DNA replication during replication itself?

Following the discovery of DNA polymerase I, what experimental evidence suggested that another enzyme is the primary enzyme of replication?

What are the features of DNA polymerase that ensure the right nucleotides are inserted?

Describe the mechanism of polymerization used by polymerases.