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Prostatic acid phosphatase

Prostatic acid phosphatase (PAP), also known as acid phosphatase 3 (ACP3) or prostatic-specific acid phosphatase (PSAP), is a non-specific phosphomonoesterase (EC 3.1.3.2) primarily synthesized and secreted by the epithelial cells of the gland. It exists as a 100 kDa dimer composed of two identical 50 kDa subunits, encoded by the ACPP located on 3q22.1, and features three N-linked sites that contribute to its stability and secretion. PAP is abundantly present in seminal plasma, where it constitutes a major component, and its expression is upregulated in response to prostate enlargement or . In its cellular form (cPAP), PAP functions as a that dephosphorylates substrates such as ErbB-2/HER-2, thereby inhibiting cell proliferation, migration, and tumorigenicity while enhancing androgen sensitivity. Extracellularly, as a secreted (sPAP), it acts as an ectonucleotidase that hydrolyzes extracellular to generate , which suppresses pain signaling by activating A1 adenosine receptors in the . Additionally, PAP hydrolyzes in seminal fluid post-ejaculation to produce free choline, potentially supporting sperm function and fertility. Its activity is optimal at acidic pH (around 5.5) and is inhibited by , distinguishing it from other acid phosphatases. Clinically, PAP has been a longstanding biomarker for prostate cancer since the 1940s, with elevated serum levels (>2.5 U/L) indicating advanced or metastatic disease and correlating with poorer prognosis, higher Gleason scores, and reduced cancer-specific survival. Although largely supplanted by (PSA) for initial screening per American Urological Association guidelines, PAP remains valuable for monitoring castration-resistant and as a prognostic indicator in intermediate- to high-risk cases. It also serves as an antigenic target in , as exemplified by (Provenge), an FDA-approved vaccine that targets PAP to extend median survival by approximately 4 months in metastatic castration-resistant patients. Beyond , PAP's amyloidogenic fragments enhance infectivity, highlighting its role in infectious disease transmission.

Biochemistry

Gene and expression

The gene encoding prostatic acid phosphatase, known by the symbols ACPP (acid phosphatase, prostate) or ACP3, is located on the long arm of human at the cytogenetic band 3q22.1, spanning genomic positions 132,317,369 to 132,368,302 on the plus strand. This positioning was confirmed through mapping studies that identified the locus within the 3q21-q23 region. The ACPP gene consists of 12 exons and produces a primary transcript that encodes a 100 kDa precursor, which undergoes post-translational modifications including N-linked . Alternative splicing of the ACPP transcript yields two main isoforms: the cellular form (cPAP), a anchored in the plasma membrane of epithelial cells, and the secreted form (sPAP), which is released extracellularly, predominantly into seminal fluid. Both isoforms are heavily glycosylated, contributing to their stability and function, though the secreted variant predominates in prostatic secretions. Expression of ACPP is highly tissue-specific, with the highest levels observed in the epithelial cells of the gland, where it constitutes a major component of seminal plasma. Lower expression occurs in other tissues, including the , , and , but these levels are significantly reduced compared to prostatic tissue. Transcriptional regulation of ACPP is primarily driven by androgens, which promote its synthesis in prostate cells, alongside influences from prostate-specific transcription factors such as that bind to promoter elements to enhance expression. This androgen-dependent control underscores its role in reproductive physiology.

Structure and enzymatic activity

Prostatic acid phosphatase (PAP), also known as ACPP or encoded by the ACPP gene, is a homodimeric glycoprotein consisting of two identical subunits, each approximately 50 kDa in molecular weight, forming a total molecular mass of about 100 kDa. The protein features three N-linked glycosylation sites at asparagine residues (Asn62, Asn188, and Asn301), which contribute to its stability and secretion. The crystal structure of human PAP, determined at 3.1 Å resolution, reveals a dimeric architecture with each subunit comprising two domains: a larger α/β domain featuring a seven-stranded mixed β-sheet flanked by α-helices, and a smaller domain composed primarily of six α-helices. In 2024, a cryo-EM structure of PAP from human semen (PDB 8XJ4) at approximately 3 Å resolution revealed an elongated homodimer conformation, differing from the compact crystal structure and indicating context-dependent assembly in physiological conditions. This overall fold is characteristic of the histidine phosphatase superfamily, with conserved structural elements facilitating catalysis. The active site of PAP is located at the interface of the two domains and includes key catalytic residues: histidine 12 (His12, acting as the nucleophile), aspartate 258 (Asp258, polarizing the P-O bond and stabilizing the intermediate), and histidine 257 (His257, serving as the general acid). Arginine 11 (Arg11) and arginine 15 (Arg15) contribute to phosphate group coordination and transition state stabilization during hydrolysis of phosphate esters. This arrangement enables a two-step mechanism involving nucleophilic attack by His12 to form a phosphohistidine intermediate, followed by hydrolysis. Classified under EC 3.1.3.2 as a non-specific , PAP exhibits optimal enzymatic activity at acidic values of 5.0–6.0. It functions as a phosphotyrosyl , dephosphorylating tyrosine-phosphorylated substrates such as ErbB-2 and phosphorylated angiotensin II with high affinity (Km in the nM range). Additionally, PAP acts as an ectonucleotidase, hydrolyzing extracellular (AMP) to , a efficient at 5.6 and requiring the transmembrane isoform for full activity in certain contexts. Kinetic studies indicate a Michaelis constant () of approximately 0.4 mM for the model substrate p-nitrophenyl phosphate (pNPP) under acidic conditions. PAP activity is notably inhibited by , with 20 mM L- reducing rates, a property that distinguishes it from tartrate-resistant acid phosphatases.

Physiological roles

Function in reproduction

Prostatic acid phosphatase (PAP), in its secreted form known as sPAcP, is abundantly present in seminal plasma at concentrations ranging from 0.3 to 1 mg/mL, making it one of the predominant glycoproteins derived from the . This enzyme constitutes a significant portion of the prostatic contribution to , where it functions primarily to facilitate the post-ejaculatory processing of seminal fluid. sPAcP exhibits the highest activity among acid phosphatases in human , with normal levels averaging around 400 U/mL, reflecting its specialized role in male reproductive . By dephosphorylating substrates like and in seminal fluid, sPAcP further supports and , essential steps for fertilization. The generation of choline from hydrolysis, in particular, promotes signaling that enhances sperm hyperactivation and progression through the female reproductive tract. These activities underscore sPAcP's contribution to fertility by optimizing the seminal environment for function. The precise physiological role of PAP in remains somewhat elusive, though recent research as of 2023 emphasizes its role in choline production for . The evolutionary conservation of PAP's across mammalian species highlights its fundamental importance for , with similar expression patterns and functions observed in various mammals.

Mechanism of pain suppression

Prostatic acid phosphatase (PAP), particularly its cellular transmembrane isoform (cPAcP), functions as an ecto-5'-nucleotidase in the , hydrolyzing extracellular (AMP) to generate . This enzymatic activity occurs at the surface of nociceptive neurons, where the produced adenosine binds to and activates A1 adenosine receptors, thereby inhibiting transmission without involving pathways. Intrathecal administration of PAP produces potent analgesia that outlasts traditional opioids, with effects persisting for up to 3 days in rodent models of inflammatory and , compared to mere hours for . This prolonged duration arises from sustained signaling, which hyperpolarizes nociceptive fibers and reduces their excitability in the dorsal horn of the . cPAcP is predominantly expressed in small-diameter (DRG) neurons and lamina II dorsal horn neurons of the , as well as in peripheral sensory nerves, positioning it to modulate nociceptive input at both peripheral and central sites. In conditions, such as models, PAP expression in DRG neurons can be modulated. Experimental evidence from PAP mice demonstrates heightened sensitivity to thermal and mechanical in models of chronic inflammatory and , underscoring the enzyme's role in adenosine-mediated relief. These mice exhibit normal acute responses but fail to generate sufficient extracellular upon AMP hydrolysis, confirming that PAP's analgesic effects are independent of receptors and rely specifically on A1 receptor activation.

Role in prostate cancer

Pathophysiological involvement

In , dysregulation of prostatic acid phosphatase (PAP), particularly its cellular isoform (cPAP), plays a significant role in tumor progression. Downregulation of cPAP expression is commonly observed in advanced stages of the disease, where it promotes androgen-independent growth of cancer cells. This occurs through increased phosphorylation of the ErbB-2 receptor (also known as HER-2), as cPAP normally functions as a phosphatase that dephosphorylates ErbB-2 at key residues such as Tyr-1221/1222, thereby inhibiting downstream signaling pathways that drive . Loss of this dephosphorylation activity allows unchecked ErbB-2 activation, enhancing tumorigenicity and resistance to . The loss of cPAP phosphatase activity further contributes to dysregulated , leading to sustained and reduced in cells. Studies have shown that prostate cancer cell lines with low cPAP levels exhibit higher growth rates and invasiveness compared to those with normal expression, underscoring the tumor-suppressive role of this enzyme. Overall, these expression changes reflect an inverse relationship between PAP activity and disease aggressiveness. Low cPAP levels serve as a prognostic indicator in , correlating with higher Gleason scores, increased risk of , and poorer overall survival. For instance, reduced cPAP expression is associated with Gleason scores of 8 or higher, which denote more aggressive tumors, and has been linked to a higher likelihood of biochemical recurrence and distant spread. This inverse correlation with tumor progression highlights cPAP's potential as a marker of severity, independent of other factors. Patients with diminished PAP expression often experience shorter -free survival intervals, emphasizing the clinical relevance of these molecular alterations. PAP expression patterns complement () in evaluating tumor aggressiveness, providing additive prognostic information without implying a direct causative role in oncogenesis. While PSA levels indicate overall tumor burden, low cPAP helps identify cases with heightened proliferative potential and androgen independence, improving risk stratification in intermediate- to high-risk patients. This combined assessment reveals heterogeneity in disease behavior beyond PSA alone.

Diagnostic applications

Prostatic acid phosphatase (PAP), also known as prostate-specific acid phosphatase (PSAP), serves as a for , with elevated levels in (sPAP) greater than 3.5 ng/mL indicating potential involvement, particularly in advanced or metastatic disease. The specificity of the assay is enhanced by measuring the tartrate-labile fraction, which isolates prostatic enzyme activity from non-prostatic sources, reducing interference from other tissues. In , sPAP is particularly useful for detecting recurrence after , where rising levels signal tumor progression or residual disease. In (IHC), PAP staining is positive in approximately 80-95% of prostate adenocarcinomas, providing a reliable marker for confirming prostatic origin in tissue samples. This staining pattern helps distinguish from non-prostatic tumors, such as urothelial , which typically show negative PSAP reactivity. Although PAP was a primary diagnostic tool before the 1980s, it was largely supplanted by (PSA) due to the latter's superior sensitivity for early detection (95% vs. 60% for PAP). Recent studies have revived interest in PAP, particularly in combination with PSA and other markers, to improve specificity for advanced where PSA alone may lack precision. Current guidelines do not recommend PAP as a routine first-line screening test, given its lower sensitivity compared to PSA, but endorse its use as an adjunct in equivocal cases or to identify the prostatic of metastases from unknown primary sites.

Role in HIV infection

Formation of SEVI amyloids

SEVI, or semen-derived enhancer of virus infection, consists of amyloid formed from proteolytic fragments of the secreted form of prostatic acid phosphatase (sPAcP) present in human . These arise through the of specific peptides derived from sPAcP, which is secreted by the prostate into seminal fluid. The formation process begins with proteolytic cleavage of sPAcP during semen liquefaction, generating peptides such as PAP<sub>248–286</sub> (a 39-amino-acid fragment) and PAP<sub>85–120</sub> (a 36-amino-acid fragment), both of which are capable of aggregating into structures. These peptides adopt beta-sheet-rich conformations and self-assemble into at neutral , as found in (approximately pH 7.2–8.0), with aggregation occurring rapidly within minutes to hours under physiological agitation. The C-terminal residues, particularly in PAP<sub>248–286</sub>, play a critical role in nucleating this fibrillization, leading to ordered, amyloidogenic beta-hairpin structures. The resulting SEVI fibrils exhibit characteristic physical properties, including widths of 5–20 nm and lengths up to several micrometers, forming positively charged structures due to their composition. These fibrils bind electrostatically to negatively charged and viral envelopes, with sPAcP concentrations in ranging from 1–2 mg/mL contributing to SEVI levels of approximately 35–40 μg/mL. studies demonstrate that isolated PAP-derived peptides, such as synthetic PAP<sub>248–286</sub>, form these amyloid fibrils independently of other seminal components, as confirmed by T fluorescence assays and .

Enhancement of viral transmission

SEVI, formed from fragments of prostatic acid phosphatase (), potently enhances HIV-1 infectivity by promoting viral attachment to target cells such as dendritic cells and macrophages. This enhancement occurs primarily through charge neutralization of the negatively charged , which overcomes electrostatic repulsions between the virions and host cell surfaces, resulting in a 10- to 100-fold increase in attachment efficiency. Studies using primary HIV-1 isolates have confirmed this boost , demonstrating SEVI's ability to amplify infection across various viral strains and cell types. The mechanism involves SEVI amyloid fibrils serving as bridges that capture and concentrate HIV virions on target cell surfaces, thereby facilitating direct contact and entry. These , derived from the self-assembly of PAP peptides, enhance viral fusion and without altering coreceptor requirements. Furthermore, SEVI promotes HIV-1 across mucosal epithelial barriers, aiding initial penetration during sexual transmission. This enhancement contributes to the epidemiological relevance of in HIV-1 , where the probability of per sexual act can reach up to 1% under high-risk conditions, underscoring SEVI's role in elevating low inocula to infectious thresholds. Observations with primary isolates from semen-exposed scenarios further support this, highlighting SEVI's impact on dynamics.

Therapeutic applications

Immunotherapy for prostate cancer

Prostatic acid phosphatase (PAP) serves as a tumor-associated in , making it a key target for strategies aimed at eliciting antitumor immune responses. The most established approach is (Provenge), an autologous cellular approved by the FDA in 2010 for the treatment of or minimally symptomatic metastatic castration-resistant (mCRPC). This is produced by isolating a patient's peripheral blood mononuclear cells, activating them with PAP fused to (GM-CSF), and reinfusing the PAP-loaded dendritic cells to prime T-cell immunity against PAP-expressing cancer cells. The therapy is administered in three infusions over approximately , with patient selection typically limited to those with advanced disease who have no visceral metastases and are not candidates for . The mechanism of involves activating antigen-presenting cells to induce a PAP-specific CD4+ and CD8+ T-cell response, which targets and lyses cells overexpressing , a protein elevated in up to 95% of tumors. Pivotal phase 3 clinical trials, including the study, demonstrated that extends median overall survival by 4.1 months (25.8 months versus 21.7 months in ) and reduces the risk of death by 22% in mCRPC patients, with no significant impact on time to disease progression but a favorable profile. Common side effects are mild and transient, including flu-like symptoms such as , fever, and , occurring in over 90% of patients but rarely leading to discontinuation. Beyond sipuleucel-T, other PAP-targeted immunotherapies are under investigation, including DNA and peptide vaccines designed to directly stimulate PAP-specific T-cell responses. For instance, the DNA vaccine pTVG-HP, which encodes PAP and GM-CSF, has shown induction of robust antigen-specific T-cell immunity in phase 1 and 2 trials for mCRPC, with ongoing phase 2 studies as of November 2025 exploring combinations to enhance efficacy; recent findings suggest robust immune responses but recommend using pTVG-HP alone in future studies due to limited added benefit from additional antigens. Additionally, checkpoint inhibitors such as PD-1/PD-L1 blockers are being combined with PAP vaccines in phase 2 and 3 trials to potentiate T-cell activation, with preliminary data from 2025 studies indicating improved response rates in advanced prostate cancer when paired with sipuleucel-T or DNA platforms. These approaches highlight the evolving role of PAP as a cornerstone antigen in prostate cancer immunotherapy.

Potential antiviral and analgesic uses

Prostatic acid phosphatase (PAP) fragments contribute to the formation of semen-derived enhancer of viral infection (SEVI) amyloids, which facilitate HIV-1 attachment to cells and enhance transmission during . To counter this, researchers have developed SEVI-neutralizing agents, including small-molecule inhibitors like surfen and , which bind to SEVI and prevent their interaction with particles, thereby reducing HIV-1 infectivity in models by up to 90-100% depending on concentration. These compounds demonstrate potential as components of topical microbicides for HIV prevention, with preclinical studies showing effective blockade of semen-mediated enhancement in cervical tissue explants. In parallel, peptides and amyloid-binding agents, such as oligovalent derivatives, have been engineered to disassemble SEVI structures or inhibit their formation from PAP precursors, restoring HIV infectivity to near-baseline levels and suggesting utility in vaginal or rectal formulations. As of November 2025, these approaches remain in , with ongoing efforts to optimize formulations for mucosal stability and compatibility with existing antiretrovirals like tenofovir for synergistic prevention strategies. Challenges in antiviral applications include the inherent of SEVI amyloids in seminal , which resist and require inhibitors with prolonged activity in protein-rich environments, as well as ensuring specificity to avoid disrupting beneficial seminal components. Future directions emphasize integrating SEVI inhibitors into combination microbicides to address transmission bottlenecks, potentially enhancing efficacy against diverse viral strains. Beyond antivirals, PAP exhibits analgesic potential through its ectonucleotidase activity, which hydrolyzes extracellular to , activating A1 adenosine receptors to suppress nociceptive signaling in the . Recombinant or PAP administered intrathecally in rodent models of inflammatory and produces dose-dependent antinociception lasting days to weeks, without evidence of tolerance or addiction associated with opioids. This mechanism supports spinal delivery routes, such as intrathecal injections, to target conditions like or , bypassing systemic exposure. Adenosine analogs, including A1 receptor agonists like NECA, mimic PAP's effects and provide rapid relief in preclinical assays, offering a non-enzymatic alternative for therapeutic development. However, off-target effects pose hurdles, as elevated can induce cardiovascular side effects like or via non-spinal receptor activation. As of November 2025, no phase I human trials for recombinant PAP analgesics have advanced, but the approach aligns with broader efforts for non-opioid therapies amid the ongoing opioid crisis, with potential for localized delivery to minimize systemic risks.

History

Discovery and initial characterization

Prostatic acid phosphatase (PAP), an with optimal activity at acidic , was first identified through studies of tissue extracts. In , researchers Wilhelm Kutscher and Heinrich Wolbergs discovered exceptionally high levels of this in human gland tissue and , distinguishing it from other phosphatases due to its substrate specificity and pH dependence. Their work laid the for recognizing PAP as a prostate-derived enzyme, with concentrations in ranging from a minimum of 400 units per ml, typically 1500–3500 units per ml, far exceeding levels in other organs. Building on this, American biochemists Alexander B. Gutman, Edith E. Sproul, and Ethel B. Gutman extended the characterization in 1936 by examining phosphatase activity in bone metastases from patients. They observed elevated " at sites of osteoplastic metastases, suggesting a link between the enzyme's production in prostatic cells and its release during progression. This finding highlighted PAP's potential as a marker for advanced prostatic , though serum levels remained unexamined at the time. The seminal advancement came in 1938 when Alexander B. Gutman and Ethel B. Gutman reported markedly elevated " levels specifically in patients with metastasizing prostatic , particularly those with involvement. In their study published in the , they measured activity using a modified and found values exceeding 10 units (Bodansky method) in over 80% of cases with metastases, while levels were normal in non-metastatic disease or other cancers. No significant elevations occurred in prostatic diseases without metastases or in women, underscoring the enzyme's prostatic origin. This work, along with follow-up reports in 1940 on clinical correlations, established as a biochemical indicator of metastatic spread. In the 1940s, further refinements confirmed PAP's prostate specificity, with Huggins and Hodges in 1941 demonstrating that androgen deprivation via or therapy reduced PAP levels in parallel with tumor , solidifying its role in treatment response. By the early 1950s, its presence in was more thoroughly linked to prostatic secretory function, and specificity was enhanced through the of tartrate inhibition; Fishman and Lerner developed an in 1953 showing that L- selectively inhibits prostatic isoenzyme activity by over 90%, allowing differentiation from non-prostatic acid phosphatases in . These developments marked the initial biochemical profiling of PAP before its broader clinical integration.

Development as a clinical tool

In the mid-20th century, prostatic acid (PAP) emerged as the first clinically viable for , with elevated levels detected in patients with advanced disease as early as the 1940s. By the 1950s and 1960s, its measurement became a standard for monitoring disease progression, staging tumors, and assessing treatment responses, particularly after therapies like or administration, due to its correlation with metastatic spread. Widespread adoption in during the 1970s solidified PAP's role in guiding therapeutic decisions, though its utility was limited to advanced cases with bone metastases. The introduction of () in the early 1980s marked a significant shift, as demonstrated superior for detecting both localized and metastatic , leading to a rapid decline in PAP's use as a marker. Despite this, PAP retained value in (IHC) for pathological diagnosis, where it serves as a reliable marker for confirming prostatic origin in tissue samples, often used alongside staining. The of the human (ACPP, also known as ACP3) in 1989 facilitated molecular studies and renewed interest in its functions beyond . In the 2000s, PAP experienced a resurgence with the discovery of its roles in modulation—acting as an ectonucleotidase to generate and suppress signals—and in transmission, where PAP-derived peptides form (SEVI) that enhance viral infectivity. Key milestones include the 2010 FDA approval of (Provenge), the first autologous cellular targeting for metastatic castration-resistant , which demonstrated a 22.5% reduction in mortality risk in phase III trials. In the , research has advanced toward leveraging SEVI inhibition for prevention, with studies identifying amyloid-binding agents that disrupt fibril formation and reduce viral enhancement in seminal fluid models. As of 2025, ongoing research has explored as a target for chimeric receptor (CAR) T-cell therapy, showing promising antitumor efficacy in preclinical models, and developed high-affinity fully human antibodies against for potential targeted cancer therapies.