Tartrate-resistant acid phosphatase

Tartrate-resistant acid phosphatase (TRAP or TRAPase), also called acid phosphatase 5, tartrate resistant (ACP5), is a is_associated_with::glycosylated is_associated_with::monomeric is_associated_with::metalloprotein is_associated_with::enzyme expressed in mammals. It has a molecular weight of approximately 35kDa, a basic is_associated_with::isoelectric point (7.6-9.5), and optimal activity in acidic conditions. TRAP is synthesized as latent is_associated_with::proenzyme and activated by is_associated_with::proteolytic cleavage and reduction. It is differentiated from other mammalian acid is_associated_with::phosphatases by its resistance to inhibition by tartrate, molecular weight and characteristic purple colour.

The mechanism of phosphate ester hydrolysis by TRAP is through a nucleophilic attack mechanism, whereby, catalysis occurs with the binding of a phosphate-substrate to the Fe2+ in the active site of TRAP. This is then followed by a nucleophilic attack by a hydroxide ligand on the bound phosphorus atom, resulting in cleavage of the phosphate ester bond and production of an alcohol. The exact identity and mechanism of the hydroxide ligand is unclear, but it is thought to be either a hydroxide that bridges the metal ions within the active site or a terminal hydroxide bound to Fe3+, with conflicting reports for both mechanisms.

TRAP expression and cell localization
Under normal circumstances, TRAP is highly expressed by is_associated_with::osteoclasts, activated is_associated_with::macrophages, is_associated_with::neurons, and by the porcine endometrium during pregnancy. In newborn rats, TRAP is also detectable in the spleen, thymus, liver, kidneys, skin, lung, and heart at low levels. TRAP expression is increased in certain pathological conditions. These include leukaemic reticuloendotheliosis (is_associated_with::hairy cell leukaemia), is_associated_with::Gaucher’s disease, is_associated_with::HIV-induced encephalopathy, is_associated_with::osteoclastoma and is_associated_with::osteoporosis, and metabolic bone diseases.

In osteoclasts, TRAP is localized within the ruffled border area, the lysosomes, the Golgi cisternae and vesicles.

TRAP gene, promoter organisation and transcription
Mammalian TRAP is encoded by one gene, which is localized on chromosome 19 (19p13.2-13.3) in humans, and on chromosome 9 in mice. TRAP DNA is, as expected from is_associated_with::protein sequencing, highly conserved throughout the class mammalia. The TRAP gene has been cloned and sequenced in porcine, rat, human, and murine species. Human, murine, and porcine TRAP genes all contain 5 exons, and have the ATG codon at the beginning of exon 2, with exon 1 being non-coding. Within the exon 1 promoter, there are three distinct “tissue-specific” promoters: 1A, 1B, and 1C. This would allow TRAP expression to be tightly controlled. Transcribed from this gene is a 1.5kb mRNA with an open reading frame (ORF) of 969-975 bp encoding a 323-325 amino acid protein. In the rat, the ORF is 981 bp in length and encodes for a 327-amino acid protein. TRAP is translated as a single polypeptide. TRAP gene transcription is regulated by the is_associated_with::Microphthalmia-associated transcription factor.

Physiology
The exact physiologic role(s) of TRAP is unknown, but many functions have been attributed to this protein. In knockout studies, TRAP−/− mice exhibit mild is_associated_with::osteopetrosis, associated with reduced osteoclast activity. These result in thickening and shortening of the cortices, formation of club-like deformities in the distal is_associated_with::femur, and widened epiphyseal growth plates with delayed mineralization of cartilage, all of which increase with age. In TRAP overexpressing transgenic mice, mild osteoporosis occurs along with increased is_associated_with::osteoblast activity and is_associated_with::bone synthesis. Proposed functions of TRAP include is_associated_with::osteopontin /is_associated_with::bone sialoprotein is_associated_with::dephosphorylation, the generation of is_associated_with::reactive oxygen species (ROS), iron transport, and as a cell growth and differentiation factor.

Protein dephosphorylation and osteoclast migration
It has been shown that osteopontin and bone sialoprotein, bone matrix phosphoproteins, are highly efficient in vitro TRAP substrates, which bind to osteoclasts when phosphorylated. Upon partial dephosphorylation, both osteopontin and bone sialoprotein are incapable of binding to is_associated_with::osteoclasts. From this effect, it has been hypothesized that TRAP is secreted from the ruffled border, dephosphorylates osteopontin and allows osteoclast migration, and further resorption to occur.

ROS generation
Reactive oxygen species (ROS) are generated in macrophages and osteoclasts from is_associated_with::superoxide (O2−.), which forms from the action of NADPH-oxidase on oxygen (O2). They play an essential role in the function of phagocytic cells.

TRAP, containing a redox active iron, catalyzes the generation of ROS through Fenton chemistry:


 * O2 → (NADPH-oxidase) O2- ∙ → (superoxide dismutase) H2O2 → (catalase) H2O + O2
 * TRAP-Fe3+ (purple) + O2- ∙→ TRAP-Fe2+ (pink) + O2
 * H2O2 + TRAP-Fe2+ (pink) → HO∙ + HO− + TRAP-Fe3+

producing is_associated_with::hydroxyl radicals, is_associated_with::hydrogen peroxide, and singlet oxygen. In osteoclasts, ROS are generated at the ruffled border and seem to be required for resorption and degradation to occur.

Iron transport
In the pregnant sow, uteroferrin is highly expressed in the uterine fluids. Due to the unique anatomy of the porcine uterus, and the specific, progesterone-induced expression of TRAP; it is hypothesized that uteroferrin acts as an iron transport protein.

Cell growth and differentiation factor
TRAP is associated with osteoblast migration to bone resorption sites, and, once there, TRAP is believed to initiate osteoblast differentiation, activation, and proliferation. This hypothesis was formed from the examination of the bone structure of TRAP-null mice. It was noted that, in addition to is_associated_with::osteopetrosis, bone formation occurred in a haphazard manner, where the microarchitecture was highly irregular.

In TRAP overexpressing mice, it has been found that the affected mice are grossly obese. This has led to the hypothesis that TRAP has involvement in hyperplastic obesity.