Carbonic anhydrase

Ribbon diagram of human carbonic anydrase II, with zinc molecule visible in the center

The carbonic anhydrases (or carbonate dehydratases) form a family of enzymes that catalyze the rapid conversion of carbon dioxide to bicarbonate and protons, a reaction that occurs rather slowly in the absence of a catalyst. The active site of most carbonic anhydrases contains a zinc ion; they are therefore classified as metalloenzymes.

Structure and function of carbonic anhydrase

Several forms of carbonic anhydrase occur in nature. In the best-studied α-carbonic anhydrase form present in animals, the zinc ion is coordinated by the imidazole rings of 3 histidine residues, His94, His96 and His119.

The primary function of the enzyme in animals is to interconvert carbon dioxide and bicarbonate to maintain acid-base balance in blood and other tissues, and to help transport carbon dioxide out of tissues.

There exist at least 14 different isoforms in mammals. Plants contain a different form called β-carbonic anhydrase, which, from an evolutionary standpoint, is a distinct enzyme, but participates in the same reaction and also uses a zinc ion in its active site. In plants, carbonic anhydrase helps raise the concentration of CO₂ within the chloroplast in order to increase the carboxylation rate of the enzyme Rubisco. This is the reaction that integrates CO₂ into organic carbon sugars during photosynthesis, and can use only the CO₂ form of carbon, not carbonic acid or bicarbonate.

In 2000, a cadmium-containing carbonic anhydrase was found to be expressed in marine diatoms during zinc limitation. In the open ocean, zinc is often in such low concentrations that it can limit the growth of phytoplankton like diatoms; thus a carbonic anhydrase using a different metal ion would be beneficial in these environments. Before this discovery, cadmium has generally been thought of as a very toxic heavy metal without biological function. As of 2005, this peculiar carbonic anhydrase form hosts the only known beneficial cadmium-dependent biological reaction.

The reaction catalyzed by carbonic anhydrase is:

${\rm {CO_{2}+H_{2}O\rightarrow ^{Carbonic\ anhydrase}HCO_{3}^{-}+H^{+}}}$(in tissues - high CO₂ concentration)

The reaction rate of carbonic anhydrase is one of the fastest of all enzymes, and its rate is typically limited by the diffusion rate of its substrates. Typical catalytic rates of the different forms of this enzyme ranging between 10⁴ and 10⁶ reactions per second.

The reverse reaction is also relatively slow (kinetics in the 15-second range), which is why a carbonated drink does not instantly degas when opening the container, but will rapidly degas in one's mouth when carbonic anhydrase is added with saliva.

${\rm {HCO_{3}^{-}+H^{+}\rightarrow H_{2}CO_{3}\rightarrow CO_{2}+H_{2}O}}$ (in lungs and nephrons of the kidney - low CO₂ concentration, in plant cells)

Mechanism

Close-up rendering of active site of human carbonic anhydrase II, showing three histidine residues (in pink) and a hydroxide group (red and white) coordinating the zinc ion (purple). From PDB: 1CA2​.

A zinc prosthetic group in the enzyme is coordinated in three positions by histidine side chains. The fourth coordination position is occupied by water. This causes polarisation of the hydrogen-oxygen bond, making the oxygen slightly more negative, thereby weakening it.

A fourth histidine is placed close to the substrate of water and accepts a proton, in an example of general acid-general base catalysis. This leaves a hydroxide attached to the zinc.

The active site also contains specificity pocket for carbon dioxide, bringing it close to the hydroxide group. This allows the electron rich hydroxide to attack the carbon dioxide, forming bicarbonate.

CA families

Ribbon diagram of human carbonic anhydrase II. Active site zinc ion visible at center. From PDB: 1CA2​.

There are at least five distinct CA families (α, β, γ, δ and ε). These families have no significant amino acid sequence similarity and in most cases are thought to be an example of convergent evolution. The α-CAs are found in humans.

α-CA

The CA enzymes found in mammals are divided into four broad subgroups, which, in turn consist of several isoforms:

• the cytosolic CAs (CA-I, CA-II, CA-III, CA-VII and CA XIII) (CA1, CA2, CA3, CA7, CA13)
• mitochondrial CAs (CA-VA and CA-VB) (CA5A, CA5B)
• secreted CAs (CA-VI) (CA6)
• membrane-associated CAs (CA-IV, CA-IX, CA-XII, CA-XIV and CA-XV) (CA4, CA9, CA12, CA14)

There are three additional "acatalytic" CA isoforms (CA-VIII, CA-X, and CA-XI) (CA8, CA10, CA11) whose functions remain unclear.

Comparison of human carbonic anhydrases

Isoform	Gene	Molecular mass	Location (cell)	Location (tissue)	Relative activity	Sensitivity to sulfonamides
CA-I	CA1	29 kDa	cytosol	RBCs and GI tract	15%	high
CA-II	CA2	29 kDa	cytosol	almost ubiquitous	100%	high
CA-III	CA3	29 kDa	cytosol	8% of soluble protein in Type I muscle	1%	low
CA-IV	CA4	35 kDa	extracellularily GPI-linked	Widely distributed, e.g. acid-transporting epithelium	~100%	moderate
CA-VA	CA5A		mitochondria
CA-VB	CA5B		mitochondria	secreting cells
CA-VI	CA6
CA-VII	CA7		cytosol
CA-IX	CA9		cell membrane-associated
CA-XII	CA12	44 kDa	extracellularily located active site	certain cancers	~30%
CA XIII	CA13		cytosol
CA-XIV	CA14	54 kDa	extracellularily located active site	kidney, heart, skeletal muscle, brain
and CA-XV

β-CA

Most prokaryotic and plant chloroplast CAs belong to the beta family. Two signature patterns for this family have been identified:

• C-[SA]-D-S-R-[LIVM]-x-[AP]
• [EQ]-[YF]-A-[LIVM]-x(2)-[LIVM]-x(4)-[LIVMF](3)-x-G-H-x(2)-C-G

γ-CA

The gamma class of CAs come from methane-producing bacteria that grow in hot springs.

δ-CA

The delta class of CAs has been described in diatoms. The distinction of this class of CA has recently come into question, however.

ε-CA

The epsilon class of CAs occurs exclusively in bacteria in a few chemolithotrophs and marine cyanobacteria that contain cso-carboxysomes. Recent 3-dimensional analyses suggest that ε-CA bears some structural resemblance to β-CA, particularly near the metal ion site. Thus, the two forms may be distantly related, even though the underlying amino acid sequence has since diverged considerably.

Pharmacological agents affecting CA

See Carbonic anhydrase inhibitors

External links

• PDB Molecule of the Month Carbonic anhydrase