A carboxypeptidase ( EC number 3.4.16 - 3.4.18) is a

protease A protease (also called a peptidase, proteinase, or proteolytic enzyme) is an enzyme that catalyzes (increases reaction rate or "speeds up") proteolysis, breaking down proteins into smaller polypeptides or single amino acids, and spurring the ...

enzyme Enzymes () are proteins that act as biological catalysts by accelerating chemical reactions. The molecules upon which enzymes may act are called substrates, and the enzyme converts the substrates into different molecules known as products ...

that hydrolyzes (cleaves) a peptide bond at the

carboxy-terminal The C-terminus (also known as the carboxyl-terminus, carboxy-terminus, C-terminal tail, C-terminal end, or COOH-terminus) is the end of an amino acid chain (protein or polypeptide), terminated by a free carboxyl group (-COOH). When the protein is ...

(C-terminal) end of a

protein Proteins are large biomolecules and macromolecules that comprise one or more long chains of amino acid residues. Proteins perform a vast array of functions within organisms, including catalysing metabolic reactions, DNA replication, res ...

peptide Peptides (, ) are short chains of amino acids linked by peptide bonds. Long chains of amino acids are called proteins. Chains of fewer than twenty amino acids are called oligopeptides, and include dipeptides, tripeptides, and tetrapeptides. ...

. This is in contrast to an

aminopeptidase Aminopeptidases are enzymes that catalyze the cleavage of amino acids from the amino terminus ( N-terminus) of proteins or peptides (exopeptidases). They are widely distributed throughout the animal and plant kingdoms and are found in many subcel ...

s, which cleave peptide bonds at the N-terminus of proteins. Humans, animals, bacteria and plants contain several types of carboxypeptidases that have diverse functions ranging from catabolism to protein maturation. At least two mechanisms have been discussed.

Functions

Initial studies on carboxypeptidases focused on pancreatic carboxypeptidases A1, A2, and B in the

digestion Digestion is the breakdown of large insoluble food molecules into small water-soluble food molecules so that they can be absorbed into the watery blood plasma. In certain organisms, these smaller substances are absorbed through the small intest ...

of food. Most carboxypeptidases are not, however, involved in catabolism. Instead they help to mature proteins, for example

Post-translational modification Post-translational modification (PTM) is the covalent and generally enzymatic modification of proteins following protein biosynthesis. This process occurs in the endoplasmic reticulum and the golgi apparatus. Proteins are synthesized by ribos ...

. They also regulate biological processes, such as the biosynthesis of

neuroendocrine Neuroendocrine cells are cells that receive neuronal input (through neurotransmitters released by nerve cells or neurosecretory cells) and, as a consequence of this input, release messenger molecules (hormones) into the blood. In this way they bri ...

peptides such as insulin requires a carboxypeptidase. Carboxypeptidases also function in

blood clotting Coagulation, also known as clotting, is the process by which blood changes from a liquid to a gel, forming a blood clot. It potentially results in hemostasis, the cessation of blood loss from a damaged vessel, followed by repair. The mechanis ...

growth factor A growth factor is a naturally occurring substance capable of stimulating cell proliferation, wound healing, and occasionally cellular differentiation. Usually it is a secreted protein or a steroid hormone. Growth factors are important for regul ...

production,

wound healing Wound healing refers to a living organism's replacement of destroyed or damaged tissue by newly produced tissue. In undamaged skin, the epidermis (surface, epithelial layer) and dermis (deeper, connective layer) form a protective barrier again ...

, reproduction, and many other processes.

Mechanism

Carboxypeptidases hydrolyze peptides at the first amide or polypeptide bond on the C-terminal end of the chain. Carboxypeptidases act by replacing the substrate water with a carbonyl (C=O) group. The carboxypeptidase A hydrolysis reaction has two mechanistic hypotheses, via a nucleophilic water and via an anhydride. In the first proposed mechanism, a promoted-water pathway is favoured as Glu270 deprotonates the nucleophilic water. The Zn²⁺ ion, along with positively charged residues, decreases the pKa of the bound water to approximately 7. Glu 270 has a dual role in this mechanism as it acts as a base to allow for the attack at the amide carbonyl group during nucleophilic addition. It acts as an acid during elimination when the water proton is transferred to the leaving nitrogen group. The oxygen on the amide carbonyl group does not coordinate to the Zn²⁺ until the addition of the water. The deprotonation of the Zn²⁺ coordinated water by Glu 270 provides an activated hydroxide nucleophile which attacks the amide carbonyl group in the peptide bond in a nucleophilic addition. The negatively charged intermediates that are formed during hydrolysis are stabilized by the Zn²⁺ ion. The interaction between the carbonyl group and the neighbouring arginine, Arg 217, also stabilizes the negatively charged intermediates. The zinc-bound hydroxide interacts with the amide with the electrostatic stabilization of the transition state provided by the Zn²⁺ ion and the neighbouring arginine. The second proposed mechanism via an anhydride has similar steps but there is a direct attack of Glu270 on the carbonyl group, and then the interaction of Glu270 on the Zn²⁺-bound amide forms an anhydride instead which can subsequently be hydrolyzed by water.

Classifications

By active site mechanism

Carboxypeptidases are usually classified into one of several families based on their active site mechanism. * Enzymes that use a metal in the active site are called "metallo-carboxypeptidases" (EC number 3.4.17). * Other carboxypeptidases that use active site serine residues are called "serine carboxypeptidases" (EC number 3.4.16). * Those that use an active site cysteine are called "cysteine carboxypeptidase" (or "

thiol In organic chemistry, a thiol (; ), or thiol derivative, is any organosulfur compound of the form , where R represents an alkyl or other organic substituent. The functional group itself is referred to as either a thiol group or a sulfhydryl gro ...

carboxypeptidases")(EC number 3.4.18). These names do not refer to the selectivity of the amino acid that is cleaved.

By substrate preference

Another classification system for carboxypeptidases refers to their substrate preference. * In this classification system, carboxypeptidases that have a stronger preference for those amino acids containing

aromatic In chemistry, aromaticity is a chemical property of cyclic ( ring-shaped), ''typically'' planar (flat) molecular structures with pi bonds in resonance (those containing delocalized electrons) that gives increased stability compared to satur ...

or branched hydrocarbon chains are called

carboxypeptidase A Carboxypeptidase A usually refers to the pancreatic exopeptidase that hydrolyzes peptide bonds of C-terminal residues with aromatic or aliphatic side-chains. Most scientists in the field now refer to this enzyme as CPA1, and to a related pancre ...

(A for aromatic/

aliphatic In organic chemistry, hydrocarbons ( compounds composed solely of carbon and hydrogen) are divided into two classes: aromatic compounds and aliphatic compounds (; G. ''aleiphar'', fat, oil). Aliphatic compounds can be saturated, like hexane, ...

). * Carboxypeptidases that cleave positively charged amino acids ( arginine, lysine) are called carboxypeptidase B (B for basic). A metallo-carboxypeptidase that cleaves a C-terminal glutamate from the peptide N-acetyl-L-aspartyl-L-glutamate is called " glutamate carboxypeptidase". A serine carboxypeptidase that cleaves the C-terminal residue from peptides containing the sequence -Pro-Xaa (Pro is proline, Xaa is any amino acid on the C-terminus of a peptide) is called " prolyl carboxypeptidase".

Activation

Some, but not all, carboxypeptidases are initially produced in an inactive form; this precursor form is referred to as a procarboxypeptidase. In the case of pancreatic carboxypeptidase A, the inactive

zymogen In biochemistry, a zymogen (), also called a proenzyme (), is an inactive precursor of an enzyme. A zymogen requires a biochemical change (such as a hydrolysis reaction revealing the active site, or changing the configuration to reveal the activ ...

form - pro-carboxypeptidase A - is converted to its active form - carboxypeptidase A - by the enzyme

trypsin Trypsin is an enzyme in the first section of the small intestine that starts the digestion of protein molecules by cutting these long chains of amino acids into smaller pieces. It is a serine protease from the PA clan superfamily, found in the d ...

. This mechanism ensures that the cells wherein pro-carboxypeptidase A is produced are not themselves digested.

References

External links

* {{Proteases Proteins Enzymes Metabolism pl:Karboksypeptydaza