A protein kinase
is a kinase enzyme
that modifies other proteins
by chemically adding phosphate
groups to them (phosphorylation
). This class of protein may further be separated into subsets as in the case of protein kinase C PKC alpha
, PKC beta, and PKC gamma, each with specific functions. Phosphorylation usually results in a functional change of the target protein (substrate
) by changing enzyme activity
, cellular location, or association with other proteins. Up to 30% of all proteins may be modified by kinase activity, and kinases are known to regulate the majority of cellular pathways, especially those involved in signal transduction
, the transmission of signals within the cell. The human genome contains about 500 protein kinase genes; they constitute about 2% of all eukaryotic
genes. Protein kinases are also found in bacteria and plants.
The chemical activity of a kinase involves removing a phosphate group from ATP and covalently attaching it to one of three amino acids that have a free hydroxyl group. Most kinases act on both serine and threonine, others act on tyrosine, and a number (dual specificity kinases) act on all three. There are also protein kinases that phosphorylate other amino acids, including histidine kinases that phosphorylate histidine residues.
Because protein kinases have profound effects on a cell, their activity is highly regulated. Kinases are turned on or off by phosphorylation (sometimes by the kinase itself - cis-phosphorylation/autophosphorylation), by binding of activator proteins or inhibitor proteins, or small molecules, or by controlling their location in the cell relative to their substrates.
Deregulated kinase activity is a frequent cause of disease, particularly cancer, where kinases regulate many aspects that control cell growth, movement and death. Drugs which inhibit specific kinases are being developed to treat several diseases, and some are currently in clinical use, including Gleevec (imatinib) and Iressa (gefitinib).
Serine/threonine-specific protein kinases
Serine/threonine protein kinases phosphorylate the OH group of serine or threonine (which have similar sidechains). Activity of these protein kinases can be regulated by specific events (e.g. DNA damage), as well as numerous chemical signals, including cAMP/cGMP, Diacylglycerol, and Ca2+/calmodulin.
One very important group of protein kinases are the MAP kinases (acronym from: "mitogen/microtubule-activated protein kinases"). Important subgroups are the kinases of the ERK subfamily, typically activated by mitogenic signals, and the stress-activated protein kinases JNK and p38.
While MAP kinases are Serine/threonine-specific, they are activated by combined phosphorylation on Serine/threonine and tyrosine residues. Activity of MAP kinases is restricted by a number of protein phosphatases, which remove the phosphate groups that are added to specific Serine or Threonine residues of the kinase and are required to maintain the kinase in an active conformation.
Two major factors influence activity of MAP kinases:
a) signals that activate transmembrane receptors (either natural ligands, or crosslinking agents) and proteins associated with them (mutations that simulate active state),
b) signals that inactivate the phosphatases that restrict a given MAP kinase. Such signals include oxidant stress.
Tyrosine-specific protein kinases
-specific protein kinases phosphorylate tyrosine amino acid residues, and are, like serine/threonine-specific kinases, used in signal transduction
. They act primarily as growth factor
receptors and in downstream signaling from growth factors ; some examples:
Receptor tyrosine kinases
These kinases consist of a transmembrane receptor
with a tyrosine kinase
domain protruding into the cytoplasm
. They play an important role in regulating cell division
, cellular differentiation
, and morphogenesis
. More than 50 receptor tyrosine kinases are known in mammals.
The extracellular domain serves as the ligand
-binding part of the molecule. It can be a separate unit that is attached to the rest of the receptor by a disulfide bond
. The same mechanism can be used to bind two receptors together to form a homo-
. The transmembrane element is a single α helix. The intracellular or cytoplasmic domain is responsible for the (highly conserved) kinase activity, as well as several regulatory functions.
Ligand binding causes two reactions:
- Dimerization of two monomeric receptor kinases or stabilization of a loose dimer. Many ligands of receptor tyrosine kinases are multivalent. Some tyrosine receptor kinases (e.g., the platelet-derived growth factor receptor) can form heterodimers with other similar but not identical kinases of the same subfamily, allowing a highly varied response to the extracellular signal.
- Trans-autophosphorylation (phosphorylation by the other kinase in the dimer) of the kinase.
The autophosphorylation causes the two subdomains of the intrinsic kinase to shift, opening the kinase domain for ATP binding. In the inactive form, the kinase subdomains are aligned so that ATP cannot reach the catalytic center of the kinase. When several amino acids suitable for phosphorylation are present in the kinase domain (e.g., the insulin-like growth factor receptor), the activity of the kinase can increase with the number of phosphorylated amino acids; in this case, the first phosphorylation is said to be a cis-autophosphorylation, switching the kinase from "off" to "standby".
The active tyrosine kinase phosphorylates specific target proteins, which are often enzymes themselves. An important target is the ras protein
Receptor-associated tyrosine kinases
Tyrosine kinases recruited to a receptor following hormone binding are receptor-associated tyrosine kinases and are involved in a number of signalling cascades, principally those involved in cytokine
signalling (but also others, including growth hormone
). One such receptor-associated tyrosine kinase is Janus kinase
(JAK), many of whose effects are mediated by STAT proteins
. (See JAK-STAT pathway.
Histidine-specific protein kinases
kinases are structurally distinct from most other protein kinases and are found mostly in prokaryotes
as part of two-component signal transduction mechanisms. A phosphate group from ATP is first added to a histidine residue within the kinase, and later transferred to an aspartate
residue on a 'receiver domain' on a different protein, or sometimes on the kinase itself. The aspartyl phosphate residue is then active in signaling.
Histidine kinases are found widely in prokaryotes, as well as in plants, fungi and eukaryotes. The pyruvate dehydrogenase family of kinases in animals is structurally related to histidine kinases, but instead phosphorylate serine residues, and probably do not use a phospho-histidine intermediate.
Aspartic acid/glutamic acid-specific protein kinases
Some kinases have mixed kinase activities. For example, MEK
(MAPKK), which is involved in the MAP kinase
cascade, is a mixed serine/threonine and tyrosine kinase.
See also Protein kinase inhibitor
Kinase assays and profilings
Drug developments for kinase inhibitors are started from kinase assays
, the lead compounds are usually profiled for specificity before moving into further tests. Many profiling services are available from fluorescent based assays to radioisotope based detections