BS2093- Review

Why does CTP inhibit ATCase?

-Although not a substrate of the enzyme, increased conc. of the ATCase inhibits the activity
of CTP.
-first committed step; unique/irreversible step – makes more sense to regulate this step
before you are no longer able to

 Binds at regulatory sub-units, away from the active site.

Enzymes exist in 2 forms: T state and R state

High affinity state (hyperbolic profile )= r state

 When ATP is added, equilibrium shifted to left

 High conc of ATP suggests there is a Pur/Pyr imbalance which stimulates biosynthesis
of Pyr to maintain balance.

Low affinity = t state ( low activity, more substrate required to get same reaction as
substrate in r state)

 CTP is added, T state is stabilised -> equilibrium shifted to right.

Activity of enzyme depends on fraction of it in each state = sigmoidal shape.

What Types of Allosteric effectors are ATP and CTP?

1. Heterotopic inhibitors: Not involved in catalytic mechanism- not a substrate

BS2093 – wk. 28 – Regulation of (protein) enzyme activity

Brief Intro: Why are proteins regulated?

1. To maintain cell homeostasis

2. Efficiency
3. Responsiveness to the environment

Ways in which enzyme activity can be regulated:

 Changes in substrate conc. -> role of isoenzymes

 Binding of small effector molecules -> allosteric regulation
 Reversible covalent modification -> phosphorylation (covalent modifications)
 Binding of regulatory proteins
 Proteolytic activation
 Controlling amount of enzyme present

Lecture 1
MB2080 CYSTIC FIBROSIS

Causes multi-organ dysfunction (i.e., GI tract, respiratory system, endocrine system,

reproductory system)

Caused by:

 Defective epithelial cells: Epithelium becomes dehydrated which results in the build-
up of ‘ sticky mucus’ in the organs affected.

As of now, no cure, instead is managed by medication -> affects quality of life.

Key : Before 1940, life expectancy of individuals diagnosed with CF was 0.

*Main Symptoms:

You can get different severities of the condition, not always severe.

Several systems affected by CF:

1. Respiratory system:
 Wheezing, coughing, shortness of breath, recurring chest infection (causes damage
to airways, persistence would cause irreversible damage), exercise intolerance

Diagram shows x-ray comparing lungs of CF

patient’s vs healthy patients.

Can see that CF patients display a lot of mucus

 2. Digestive System:
 Poor weight gain and growth, diarrhoea, constipation, foul smelling greasy stools, a
bowel construction (especially in new-borns), damaged pancreas

 Mucus inhibits epithelial cells from absorbing necessary nutrients

3. Reproductive System:
 Male infertility

*Clinical manifestations

 Multifactorial complications :

*Epidemiology of Cystic Fibrosis:

- Known to be the most common life-shortening disease in white people of north

European ancestry (1 in 2500 who are born with it)
 - Small predominance of males ( at 52.4%) , and women at 47.6%.

Re-watch this part of the lecture**

Pathogenesis of Cystic fibrosis

CF is an autosomal recessive disease affecting the CFTR gene ( Cystic fibrosis

transmembrane regulator).

 The CFTR protein is an anion transporter which regulates mucus homeostasis

(hydration and ionic transport across the epithelium)

- Over 2000 variants of the CFTR gene have been identified.

- In the UK, 1 in 25 people are carriers of the defective gene (heterozygotes)

- You don’t need to have the same mutation on both chromosomes to get CF.

 Different variants and variant combinations ^ are associated with varying symptoms
of the disease.

CFTR gene and protein

CFTR gene is located on the long arm of the chromosome 7 ( Chr 7)

- The gene contains 27 exons and spans approx. 190kb of genomic DNA.
- The mRNA is 6.2 kb long (including the untranslated regions)
How is the protein activated?

 The ligand binds to its receptor that is coupled to Galpha s sub-unit. As a result,
cAMP is generated

Classification:
- 6 classifications of the disease. Each individual diagnosed fits into a category

Class I MUTATION – no protein on surface, protein production not present.

 Frame shift, splicing
This is the same with class II

 See diagram on slide ( red arrow are highly populated in diagnosed individuals)

Slide explains why variation in symptoms exists between individuals diagnosed with CF.