Cystic Fibrosis

The cystic fibrosis transmembrane conductance regulator

(CFTR)
• The CFTR protein is made up of 1,480
amino acids. Once the CFTR protein
chain is made, it is folded into a
specific 3-D shape.
• The CFTR protein is shaped like a tube
that goes through the membrane
surrounding the cell, like a straw goes
through the plastic top on a cup.
• It is a member of the ATP binding
cassette (ABC) superfamily of proteins
which includes several clinically
important proteins
Structure of CFTR
• The Nuclear Binding Domains
• The Regulatory Domain.
• Extracellular Domains
• Transmembrane Domains
The Nuclear Binding Domains.
• Many of the mutations identified in CF occur in the first nucleotide
binding domain (NBD1), while very few occur in NBD2.
• This is a common feature of the ABC superfamily and indicates a
separate role for the two binding domains. The most common
mutation in CF, deltaF508 occurs in NBD1. This results in a 3 bp
deletion and the loss of a phenylalanine residue. The deletion causes
a protein trafficking defect. If this defect is overcome then the protein
can form a functional channel.
The Regulatory Domain
• The R domain of CFTR is encoded by exon 13 and it spans the region between NBF1 and
the second transmembrane region.
• It contains several potential sites for phosphorylation by cAMP-dependent PKA or PKC.
The activity of CFTR as an ion channel depends upon phosphorylation of the R-domain
and binding of ATP to the nuclear binding domains.
• The N terminal portion of the R-domain (RD1) is highly conserved between species but
there is a lower degree of conservation between the rest of the domain (RD2).
Extracellular Domains
• A mere 4 % of the CFTR protein is found in the extracellular loops (see
the gene sequence and structure section).
• The loops are designated according to the membrane spanning
regions they connect, M1-M2, M3-M4, M5-M6, M7-M8, M9-M10 and
M11-M12 (always odd to even).
• With the exception of the M1-M2 and the M7-M8 these extracellular
domains are very short. The M7-M8 loop contains two N-linked
glycosylation sites.
Transmembrane Domains.
• 19 % of the CFTR protein makes up the twelve transmembrane
domains (M1 - M12). These domains have been shown to be
comprised of typical a-helical secondary structure.
• Many of the residues within these regions form the channel lining
residues and have a major role in the regulation of pore function
The Intracellular Domains
• Sequences within the intracellular loops (ICL1 - 4) have been shown
to be important for the processing of CFTR and correct delivery to the
cell membrane.
• Site directed mutagenesis studies in ICL2 and ICL3 have indicated that
these sections may be close to the intracellular opening of the CFTR
pore. Changes in these regions have been shown to alter the
conductance state of the channel.
Normal and abnormal CFTR proteins. Computer illustration of a normally functioning cystic fibrosis transmembrane
conductance regulator (CFTR) protein (left) and a malfunctioning mutant CFTR protein (right) in a cell membrane (red).
Carbohydrate is yellow, chloride is green, phosphate is turquoise and adenosine triphosphate (ATP) is purple. CFTR is an ion-
channel that moves chloride and thiocyanate ions across epithelial cell membranes. Functional irregularities of these
proteins, caused by mutations of the CFTR gene, lead to malfunctioning of epithelial fluid transport, causing mucous (beige,
upper right) to build up outside the cells in the lung, pancreas and other organs, resulting in cystic fibrosis.
What Does the CFTR Protein Do?
• The CFTR protein is a particular type of protein called an ion channel.
An ion channel moves atoms or molecules that have an electrical
charge from inside the cell to outside, or from outside the cell to
inside.
• In the lung, the CFTR ion channel moves chloride ions from inside the
cell to outside the cell. To get out of the cell, the chloride ions move
through the center of the tube formed by the CFTR protein.
• Once the chloride ions are outside the cell, they attract a layer of
water. This water layer is important because it allows tiny hairs on the
surface of the lung cells, called cilia, to sweep back and forth. This
sweeping motion moves mucus up and out of the airways.
How Do Problems With the CFTR Protein
Cause CF?
• In people with CF, mutations in the CFTR gene can cause the following
problems with the CFTR protein:

• It doesn't work well

• It isn't produced in sufficient quantities
• It is not produced at all
• When any of these problems occur, the chloride ions are trapped
inside the cell, and water is no longer attracted to the space outside
the cell. When there is less water outside the cells, the mucus in the
airways becomes dehydrated and thickens, causing it to flatten the
cilia. The cilia can't sweep properly when thick, sticky mucus weighs
them down.

• Because the cilia can't move properly, mucus gets stuck in the airways,
making it difficult to breathe. In addition, germs caught in the mucus
are no longer expelled from the airway, allowing them to multiply and
cause infections.