Serine Protease Inhibitor Gene Research Articles

In Silico Genome-wide Identification and Expression Analysis of the Pin-2 Serine Protease Inhibitor Gene Family in Eggplant (Solanum melongena L.)

In Silico Genome-wide Identification and Expression Analysis of the Pin-2 Serine Protease Inhibitor Gene Family in Eggplant (Solanum melongena L.)

Identification and functional analysis of serine protease inhibitor gene family of Eocanthecona furcellata (Wolff).

The predatory natural enemy Eocanthecona furcellata plays a crucial role in agricultural ecosystems due to its effective pest control measures and defensive venom. Predator venom contains serine protease inhibitors (SPIs), which are the primary regulators of serine protease activity and play key roles in digestion, development, innate immunity, and other physiological regulatory processes. However, the regulation mechanism of SPIs in the salivary glands of predatory natural enemies is still unknown. In this study, we sequenced the transcriptome of E. furcellata salivary gland and identified 38 SPIs genes named EfSPI1∼EfSPI38. Through gene structure, multiple sequence alignment and phylogenetic tree analysis, real-time quantitative PCR (RT-PCR) expression profiles of different developmental stages and different tissues were analyzed. RNAi technology was used to explore the gene function of EFSPI20. The results showed that these 38 EfSPIs genes contained 8 SPI domains, which were serpin, TIL, Kunitz, Kazal, Antistasin, Pacifastin, WAP and A2M. The expression profile results showed that the expression of different types of EfSPIs genes was different at different developmental stages and different tissues. Most of the EfSPIs genes were highly expressed in the egg stage. The EfSPI20, EfSPI21, EfSPI22, and EfSPI24 genes of the Pacifastin subfamily and the EfSPI35 gene of the A2M subfamily were highly expressed in the nymphal and adult stages, which was consistent with the RT-qPCR verification results. These five genes are positively correlated with each other and have a synergistic effect on E. furcellata, and they were highly expressed in salivary glands. After interfering with the expression of the EfSPI20 gene, the survival rate and predatory amount of male and female adults were significantly decreased. Taken together, we speculated some EfSPIs may inhibit trypsin, chymotrypsin, and elastase, and some EfSPIs may be involved in autoimmune responses. EfSPI20 was essential for the predation and digestion of E. furcellata, and the functions of other EfSPIs were discussed. Our findings provide valuable insights into the diversity of EfSPIs in E. furcellata and the potential functions of regulating their predation, digestion and innate immunity, which may be of great significance for developing new pest control strategies.

Serine protease inhibitors LmSPN2 and LmSPN3 co-regulate embryonic diapause in Locusta migratoria manilensis (Meyen) via the Toll pathway

Female adults of the migratory locust, Locusta migratoria manilensis (Meyen), can sense seasonal photoperiod changes, which induces embryonic diapause as a key strategy to overwinter. Serine protease inhibitor genes (SPNs) were thought to play key roles during diapause, while few SPNs were functionally characterized. LmSPN2 was one of those genes differentially expressed between diapause and non-diapause eggs; however, its biological function remained to be explored. So, we conducted RNAi knockdown of LmSPN2, resulting in a significant decrease of the egg diapause rate by 29.7%. Using yeast two-hybrid assays, co-immunoprecipitation, and pull-down methods, we found an interaction between LmSPN2 and LmSPN3, which was proved to be mediated by a glutamate (E331) binding site of LmSPN2. RNAi knockdown of LmSPN3 resulted in a significant increase in diapause rate by 14.6%, indicating an inverse function of LmSPN2 and LmSPN3 on diapause regulation. Double knockdown of two SPN genes resulted in a 26.4% reduction in diapause rate, indicating that LmSPN2 was the dominant regulatory signal. Moreover, we found four Toll pathway genes (easter, spätzle, pelle, and dorsal) upregulated significantly after the knockdown of LmSPN2 while downregulated after the knockdown of LmSPN3. Therefore, we speculate that two SPNs regulate diapause through the Toll pathway. Our results indicated that LmSPN2 positively regulates locust egg entry into diapause, while LmSPN3 is a negative regulator of embryonic commitment to diapause. Their interaction is mediated by the binding site of E331 and influences egg diapause through the Toll pathway. This mechanistic understanding of diapause regulation expands our understanding of insect developmental regulation and provides functional targets for developing locust management strategies.

National survey on clinical and genetic characteristics of patients with hereditary angioedema in Latvia

BackgroundHereditary angioedema (HAE) is a rare and life-threatening inborn error of immunity. HAE is mostly caused by pathogenic variations in the serine protease inhibitor gene 1 (SERPING1), leading to deficient or dysfunctional C1-inhibitor (C1-INH), overproduction of bradykinin, and development of recurrent subcutaneous and/or submucosal oedema. The prevalence of HAE is 1 in 50,000 − 100000 people worldwide. We aimed to describe the clinical features and genetic spectrum of hereditary angioedema with C1-INH deficiency (C1-INH-HAE) in Latvia.MethodsAll patients from Latvia diagnosed with HAE (types I/II) from 2006 to March 2022 were included in the study. Laboratory tests and clinical data were analysed, and genetic tests with Sanger sequencing and whole genome sequencing were performed.ResultsThe study identified 10 C1-INH-HAE patients (nine females, one male) from eight families. The point prevalence of HAE in Latvia is 0.53 per 100 000 inhabitants. Of all patients, seven (70%) had HAE type I and three (30%) had HAE type II. The median age of patients was 54 years and the median age at onset of symptoms was 15 years. A significant delay (median 20.5 years) until diagnosis was observed, and 60% of patients had a positive family history of angioedema. All HAE patients have been hospitalised a median two times during their lifetime. Skin (100%), abdominal (80%), and airway (80%) oedema were the most frequent symptoms. Triggering factors (60%) and prodromal symptoms (90%) were referred. Attacks were severe in 50% of patients, moderate in 10%, and mild in 40%. Pathogenic variations of SERPING1 were identified in eight patients (six families), confirming the diagnosis molecularly. In two patients (two families), no pathogenic variations in the genes were found even after whole genome sequencing.ConclusionsCurrent data shows a significant delay and clear underdiagnosis of HAE in Latvia. Higher awareness and better information and communication between doctors would improve the diagnosis and management of HAE; as would screening of family members, patients with recurrent angioedema unresponsive to antihistamines and glucocorticoids, and patients with recurrent episodes of severe, unexplained abdominal pain.

Genome-wide identification and immune response analysis of serine protease inhibitor genes in the blood clam Tegillarca granosa

Serine protease inhibitors (SPIs) are the main regulators of serine protease activities. In this study, we present a genome-wide identification of SPI genes in T. granosa（TgSPI genes）and their expression characteristics in respond to Vibrio stress. A total of 102 TgSPI genes belonging to eight families, including Serpin, TIL (trypsin inhibitor like cysteine rich domain), Kunitz, Kazal, I84, Pacifastin, WAP (whey acidic protein) and A2M (Alpha-2-macroglobulin) were identified, while no genes belonging to Bowman-Birk, amfpi and Antistasin families were identified. The Kazal family has the most TgSPI genes with 38, and 11 TgSPI genes belong to the mollusc-specific I84 family. The TgSPI genes were found to be randomly distributed on 17 chromosomes with 12 tandem duplicate gene pairs. Expression profiles showed that most TgSPI genes were mainly expressed in immune-related tissues such as hepatopancreas, gill and mantle. In the hepatopancreas, most of TgSPI genes were sensitive to Vibrio stress, 28 and 29 TgSPI genes were up-regulated and down-regulated, respectively. Some up-regulated genes with signal peptides, such as the TgSPIs of I84 family, may act as a mechanism to directly prevent Vibrio from invasion. Six Kazal-type TgSPIs (TgSPI29, 45, 49, 50, 51 and 52) were intracellular proteins and their expression was down-regulated in hemocytes after Vibrio stress. This may have boosted protease activity in hemocytes to the point that more hemoglobin derived peptides were produced and secreted into the hemolymph to exert their anti-Vibrio effects. These findings may provide valuable information for further clarifying the roles of SPIs in the immune defense and will benefit future exploration of the immune function of SPIs in molluscs.

Co-expression of Cocculus hirsutus trypsin inhibitor with Cry protein reduces resistant development in targeted insects along with complete mortality

Crop losses due to various insects are a global problem. For decades, several types of research have been ongoing on this issue with limited success due to co-evolving insect pests. In this study, the modified Cocculus hirsutus trypsin inhibitor (ChTI), a serine proteinase inhibitor gene encoding a natural plant defense protein, inhibiting the digestive proteases in the midgut of phytophagous insects, was expressed alone and pyramided with the Bt-toxin Cry1Ab, to reduce resistance development against two herbivorous insects, Helicoverpa armigera and Spodoptera litura with complete insect mortality. In-silico analysis revealed the structural and conformational stability of ChTI protein. Furthermore, 80 % weight loss and reduced larval growth were observed in both insects challenged with 200 µg of the purified ChTI protein. The ChTI protein trypsin inhibitor activity was observed up to 5.5 U/mg expressed in E. coli, 3.0–3.5 U/mg in transgenic Arabidopsis plants, and up to 2.5–3.0 U/mg in transgenic tomato plants. Transgenic plants (Arabidopsis and tomato) were developed by pyramiding ChTI, and cry1Ab genes showed 100 % mortality against both insects while the transgenic plants expressing the ChTI gene alone showed up to 70 % mortality without yield penalty. Transcriptomic analysis of the transgenic plants further revealed that other pathogenesis-related defense proteins were also differently regulated.

Enhanced expression of serine protease inhibitor StSPI128 alleviates the enzymatic browning in fresh-cut potatoes via increasing antioxidant abilities

Enzymatic browning is problematic for the fresh-processed potato (Solanum tuberosum L.) industry. Previously, we identified a serine protease inhibitor gene StSPI128, which may alleviate potato browning. Therefore, the effects of StSPI128 on potato browning and browning-related factors were investigated by overexpressing the gene in this study. The results showed that overexpression of StSPI128 significantly alleviated potato browning and increased phenolic content. However, not all the decreases in protease activities or accumulations of total free amino acid and tyrosine caused by StSPI128-overexpression were significant among three transgenic lines. Besides, overexpression of StSPI128 reduced PPO activity in potatoes. Moreover, overexpression of StSPI128 increased the total antioxidant abilities and membrane stability, elevated SOD and CAT activities, and decreased ROS and MDA accumulation. In vitro experiments demonstrated that the variation in browning-substrates and PPO activities did not induce the differences in potato browning. These results indicated that enhanced expression of StSPI128 alleviated potato browning mainly by increasing antioxidant ability, not via altering browning substrates or PPO activities. This work suggested that StSPI128 is a promising potential target for browning control in fresh-processed potatoes.

A TIL-Type Serine Protease Inhibitor Involved in Humoral Immune Response of Asian Corn Borer Ostrinia furnaculis.

To elucidate the application value of insect endogenous protease and its inhibitor genes in pest control, we analyzed in detail the transcriptome sequence of the Asian corn borer, Ostrinia furnacalis. We obtained 12 protease genes and 11 protease inhibitor genes, and comprehensively analyzed of their spatiotemporal expression by qRT-PCR. In which, a previous unstudied serine protease inhibitor gene attracted our attention. It belongs to the canonical serine proteinase inhibitor family, a trypsin inhibitor-like cysteine-rich domain (TIL)-type protease inhibitor, but its TIL domain lacks two cysteine residues, and it was named as ACB-TIL. Its expression level is relatively very low in the absence of pathogen stimulation, and can be up-regulated expression induced by Gram-negative bacteria (Escherichia coli), virus (BmNPV), and dsRNA (dsEGFP), but cannot be induced by fungus spores (Metarrhizium anisopliae). Prokaryotic expressed ACB-TIL protein can significantly inhibit the melanization in vitro. Injecting this protein into insect body can inhibit the production of antimicrobial peptides of attacin, lebocin and gloverin. Inhibition of ACB-TIL by RNAi can cause the responses of other immune-, protease- and inhibitor-related genes. ACB-TIL is primarily involved in Asian corn borer humoral immunity in responses to Gram-negative bacteria and viruses. This gene can be a potential target for pest control since this will mainly affect insect immune response.

Short-term warming inhibits polyphenol oxidase activity and influences free amino acid accumulation and de novo synthesis of tyrosine in fresh-cut potato

Potatoes cut immediately after being taken out from cold storage easily undergo browning. Researchers have reported that long-term warming alleviates the browning of fresh-cut potatoes. However, long-term warming is not commercially applicable. The present study investigated the effect of short-term warming (SW) at 25 °C for 12 h on the browning of fresh-cut potatoes. The browning index of potatoes stored at 4 °C for 8 months was tested. The analysis revealed that SW efficiently blocked enzymatic browning by inhibiting polyphenol oxidase (PPO) activity, decreasing total phenolic content and malondialdehyde (MDA) content, and increasing peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT) activity, and 1,1-Diphenyl-2-picrylhydrazyl (DPPH) and 2,20-azino-bis (3-ethylben zothiazoline) 6-sulphonic acid (ABTS) inhibition rates. Moreover, SW treatment induced the expression of the serine protease inhibitor (SPI) gene and enhanced SPI activity, increased the soluble protein content and decreased free tyrosine, and lysine content in fresh-cut potatoes during the 5 d storage at 2∼4 °C. The contents of glutamic acid, aspartic acid, and proline in the fresh-cut potatoes increased after SW treatment. Furthermore, the amino acid complementation experiments showed that tyrosine and lysine aggravated browning while glutamic acid and aspartic acid reduced browning. The SW treatment reduced the activity of chorismate mutase (CM), repressed the transcript levels of CM synthesis-related genes (CM1 and CM2), and inhibited free tyrosine synthesis. These results indicate that SW treatment alleviates the browning of potato tubers after cold storage probably due to its influence on PPO, metabolism of tyrosine, other FAAs and phenolics, and antioxidants.

Mice heterozygous for the Serpinb6a null mutation show deficits in central auditory function after acoustic trauma.

Complete deficiency of the serine protease inhibitor gene, SERPINB6, is responsible for autosomal-recessive, nonsyndromic sensorineural hearing loss in humans. A mouse model of this deafness gene identifies Serpinb6a expression in the neurosensory epithelium and fibrocytes of the cochlea. Homozygous Serpinb6a mutant mice display an exaggerated hearing loss after exposure to moderate acoustic trauma. It is unknown if and how heterozygous Serpinb6a mice show increased vulnerability to acoustic trauma. We exposed Serpinb6a+/- and Serpinb6a+/+ mice to acoustic trauma and measured their hearing function prior to, 3 and 14 days postexposure, analysing shifts in hearing threshold and amplitudes of Wave I and II of the auditory brainstem-evoked response (ABR) to 4, 8, 16 and 32 kHz tones. Shifts in hearing threshold and Wave I amplitude of Serpinb6a+/- mice were not significantly different from Serpinb6a+/+ mice at both time points and all frequencies tested (P > 0.05, Mann-Whitney test). However, Wave II amplitudes at 16 and 32 kHz tones, were more severely diminished in Serpinb6a+/- mice (P < 0.05). To exclude any effects of ageing on auditory function in Serpinb6a+/- mice, hearing function of unexposed Serpinb6a+/- mice was measured at start and end of the experimental period. The shift in Wave II amplitude of exposed Serpinb6a+/- mice was significantly lower than unexposed Serpinb6a+/- mice only at 16 and 32 kHz (P < 0.01), confirming acoustic trauma as the main cause of hearing deficits in Serpinb6a+/- mice. These results suggest that heterozygous Serpinb6a humans may be vulnerable to noise.

Molecular identification, characterization, and expression analysis of a serine protease inhibitor gene from cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae).

Serine protease inhibitors (serpins), a superfamily of protease inhibitors, are known to be involved in several physiological processes, such as development, metamorphosis, and innate immunity. In our study, a full-length serpin cDNA, designated Haserpin1, was isolated from the cotton bollworm Helicoverpa armigera. The cDNA sequence of Haserpin1 is 1176 nt long, with an open reading frame encoding 391 amino acids; there is one exon and no intron. The predicted molecular weight of Haserpin1 is 43.53 kDa, with an isoelectric point of 4.98. InterProScan was employed for Haserpin1 functional characterization, which revealed that Haserpin1 contains highly conserved signature motifs, including a reactive center loop (RCL) with a hinge region (E341-N350), the serpin signature, (F367-F375) and a predicted P1-P1' cleavage site (L357-S358), which are useful for identifying serpins. Transcripts of Haserpin1 were constitutively expressed in the fat body, suggesting that it is the major site for serpin synthesis. During the developmental stages, a fluctuation in the expression level of Haserpin1 was observed, with low expression detected at the 5th-instar larval stage. In contrast, relatively high expression was detected at the prepupal stage, suggesting that Haserpin1 might play a critical role at the H. armigera wandering stage. Although the detailed function of this serpin (Haserpin1) needs to be elucidated, our study provides a perspective for the functional investigation of serine protease inhibitor genes.

Genetic Abnormalities in Pancreatitis: An Update on Diagnosis, Clinical Features, and Treatment.

Several pancreatitis susceptibility genes have been identified to date. A relationship between a mutation in the cationic trypsinogen (protease serine 1, PRSS1) gene and hereditary pancreatitis (HP) was first identified in 1996. Currently, HP has been defined as either two or more individuals within a family exhibiting pancreatitis for two or more generations, or pancreatitis linked to mutation of the PRSS1 gene. In 2000, a mutation in the serine protease inhibitor gene (Kazal type 1: SPINK1) was reported to be related to sporadic pancreatitis of unknown etiology. This paper reviews and summarizes the current published data on the pancreatitis susceptibility genes, mainly PRSS1 and SPINK1 genes, and introduces a diagnostic and therapeutic approach for dealing with patients with these gene mutations. Patients with these genetic predispositions, both children and adults, have often been initially diagnosed with idiopathic acute pancreatitis, in approximately 20–50% of pediatric cases and 28–80% of adult cases. In such patients, where the etiology is unknown, genetic testing, which requires pre-test and post-test genetic counselling, may prove helpful. Patients with chronic pancreatitis (CP) due to SPINK1 gene mutation and HP patients have a potentially high risk of pancreatic exocrine insufficiency, diabetes mellitus, and, of particular importance, pancreatic cancer. Thus, these patients require careful long-term follow-up and management. Specifically, symptomatic CP patients often need endoscopic therapy or surgery, often following a step-up approach beginning with endoscopic therapy and progressing to surgery if necessary, which is similar to the therapeutic approach for patients with CP due to other etiologies. It is important that clinicians are aware of the characteristics of patients with pancreatitis susceptibility genetic abnormalities.

Serpins: Genome-Wide Characterisation and Expression Analysis of the Serine Protease Inhibitor Family in Triticum aestivum.

The serine protease inhibitor (serpin) gene family is the largest family of protease inhibitors. Serine protease inhibitors have an active, but under-characterized, role in grain development and defense against pathogen attack in cereal crops. By exploiting publicly available genomic, transcriptomic and proteomic data for wheat (Triticum aestivum), we have identified and annotated the entire ’serpinome’ of wheat and constructed a high-quality and robust phylogenetic tree of the gene family, identifying paralogous and homeologous clades from the hexaploid wheat genome, including the Serpin-Z group that have been well characterized in barley. Using publicly available RNAseq data (http://www.wheat-expression.com/), expression profiles of the wheat serpins were explored across a variety of tissues from the developing grain, spikelet and spike. We show that the SERPIN-Z clade, among others, are highly expressed during grain development, and that there is homeologous and paralogous functional redundancy in this gene family. Further to their role in grain development, serpins play an important but under-explored role in response to fungal pathogens. Using 13 RNAseq datasets of wheat tissues infected by fungal pathogens, we identified 37 serpins with a significant disease response. The majority of the disease-responsive serpins were upregulated by Fusarium graminearum, a destructive fungal pathogen that attacks the spike and developing grain of wheat. As serpins are ubiquitous in wheat grain, the genes encoding serpins may be linked to grain development, with their disease response a result of pleiotropy.

Cloning, expression and functional verification of a serine protease inhibitor gene from Anatolica polita

Cloning, expression and functional verification of a serine protease inhibitor gene from Anatolica polita

Amelioration of Alpha-1 Antitrypsin Deficiency Diseases with Genome Editing in Transgenic Mice.

Alpha-1 antitrypsin deficiency (AATD) is a hereditary liver disease caused by mutations in the SERPINA1 serine protease inhibitor gene. Most severe patients are homozygous for PiZ alleles (PiZZ; amino acid E324K), which lead to protein aggregates in hepatocytes and reduced circulating levels of AAT. The liver aggregates typically lead to fibrosis, cirrhosis, and hepatocellular carcinoma, and the reduced circulating AAT levels can lead to emphysema and chronic obstructive pulmonary diseases. In this study, two CRISPR/Cas9 gene editing approaches were used to decrease liver aggregates and increase systemic AAT-M levels in the PiZ transgenic mouse. In the first approach, AAT expression in hepatocytes was reduced more than 98% following the systemic delivery of AAV8-CRISPR targeting exon 2 of hSERPINA1, leading to reduced aggregates in hepatocytes. In the second approach, a second adeno-associated virus, which provided the donor template to correct the Z mutation, was also administered. These treated mice had reduced AAT expression (> 98%) and a low level (5%) of wildtype AAT-M mRNA. Taken together, this study shows that CRISPR gene editing can efficiently reduce liver expression of AAT-Z and restore modest levels of wildtype AAT-M in a mouse model of AATD, raising the possibility of CRISPR gene editing therapeutic for AATD.

Host Response whenPerkinsus marinusInfection Intensities Increase in the OysterCrassostrea corteziensis

The protozoan Perkinsus marinus was identified for the first time in the oyster Crassostrea corteziensis from the state of Baja California Sur, Mexico. Intensities of P. marinus infections increased from 0.5–2 to 2–5 (Mackin scale) when water temperature was experimentally increased to 26°C for 15 days. Although disease-induced mortalities have not been reported in natural populations of C. corteziensis, high injected doses of P. marinus caused mortalities among experimental oysters. To evaluate the response of C. corteziensis with increasing levels of P. marinus, expression levels were investigated for genes involved in phagocytosis, cellular proliferation, apoptosis, and inhibitors of serine proteases. Oysters with low infection showed higher gene expression of the serine protease inhibitor gene, as well as of genes involved in phagocytosis (nicotinamide adenine dinucleotide phosphate-oxidase, actin, superoxide dismutase, and glutathione s-transferase), and genes involved in apoptosis and DNA fragmentation (tumor necrosis factor and caspase-8). Results of observed mortalities associated with high intensities of P. marinus infection indicated that C. corteziensis is a susceptible host to this pathogen. Phagocytosis and cell death by apoptosis were higher in less-infected oysters, as an apparent result of the increased expression of apoptosis-related genes.

Abstract 5516: Regulation of TFPI-2 in the progression of ovarian cancer

Abstract Ovarian cancer metastasis occurs when malignant cells migrate from the primary tumor into the ascitic fluid and form a secondary tumor in the peritoneal cavity. The cells responsible for this process, Metastases Initiating Cells (MICs), suffer a series of changes in protein expression, with increased Epithelial Mesenquimal Transition and stem cell markers having been reported. Tissue Factor Pathway Inibitor-2 (TFPI-2) is a serine proteinase inhibitor and potential tumor suppressor gene, whose reduction is correlated with more metastases and poor prognosis. As the exact stage of TFPI-2 loss is still unknown, the purpose of our study was to determine TFPI-2 during ovarian cancer progression. Protein and RNA levels were assessed in primary tumor and metastatic tissue from papillary serous carcinoma by immunohistochemistry and qPCR, respectively. TFPI-2 RNA and protein levels were analyzed in cancer cell lines and primary cultured cancer cells isolated from ascitic fluid of advanced ovarian cancer patients and in cancer spheres derived from these cultures. Epigenetic changes of 3 CpG were analized by bisulfite and pirosequencing technique in ovarian cancer cell lines and in cancer spheres derived from these cultures. We report that significantly decreased TFPI-2 protein levels in metastatic tissue compared to the primary tumor, a result confirmed in matched primary and metastatic lesions from the same ovarian cancer patients. Furthermore, cancer spheres (representing potential MICs) displayed significantly lower levels of TFPI-2 than corresponding cell line and some primary cultured cancer cells. The cancer spheres showed a modest increase in methylation percentage in three specific CpG sites within the TFPI-2 promoter. Our results suggest that TFPI-2 loss plays a role in the cancer cell escape from the primary tumor. Future studies will determine the mechanism and the biological consequences of the progressive loss of TFPI-2 in cancer. Funding: FONDECYT 11140657, 1160800, 1140970, 3140335, 3150028, CORFO L2 13IDL2-18608, IMII-P09/016-F, CONICYT-FONDAP 15130011, BMRC 13CTI21526-P6 Citation Format: Jacqueline Fry, Sumie Kato, Lorena Abarzua, Pamela Gonzalez, Carolina Ramirez, Elisa Cumsille, Erasmo Bravo, Raimundo Correa, Juan E. Leiva, Cesar Paredes, Juan C. Roa, Carolina Ibañez, Mauricio Cuello, Gareth I. Owen, Maria L. Bravo. Regulation of TFPI-2 in the progression of ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5516. doi:10.1158/1538-7445.AM2017-5516

Isolation, expression, and characterization of the serine protease inhibitor gene (600Hbpi) from Hevea brasiliensis leaves, RRIM600 cultivar

Isolation, expression, and characterization of the serine protease inhibitor gene (600Hbpi) from Hevea brasiliensis leaves, RRIM600 cultivar

Effect of ApoA4 on SERPINA3 mediated by nuclear receptors NR4A1 and NR1D1 in hepatocytes

ApoA4 exerts anti-inflammatory effects, but the mechanism remains unclear. SERPINA3 is a member of the serine proteinase inhibitor gene family, and has been shown to be involved in anti-inflammation and associated with a number of human diseases. In this study, we revealed that ApoA4 stimulates the gene expression of SERPINA3 in mouse hepatocytes both in vivo and in vitro, in a dose- and time-dependent manner. The transcriptional response of SERPINA3 to ApoA4 is regulated through the binding of ApoA4 with nuclear receptors NR4A1 and NR1D1 on the SERPINA3 promoter, which was verified with ChIP, Luciferase activity assay and RNA interference-mediated NR4A1 or NR1D1 gene knockdown. These data suggests that ApoA4 transcriptionally induced SERPINA3 expression via NR1D1 and NR4A1. Our findings may throw light on the function of ApoA4 in inflammatory responses and acute-phase reactions, as well as the development of SERPINA3 relative diseases.

The putative serine protease inhibitor (serpin) genes encoded on Echinococcus multilocularis genome and their expressions in metacestodal stage

Two putative serpin genes were identified in Echinococcus multilocularis, in addition to the already reported serpinEmu, and were designated as serpin2Emu and serpin3Emu. Western blot analysis using polyclonal antibodies against serpinEmu, putative serpin2Emu protein, and putative serpin3Emu protein indicated that all three proteins were localized in both intracellular and excretory-secretory (ES) fractions of E. multilocularis metacestodes. In addition, immune staining of parasite tissue indicated that all three proteins were localized at the germinal layer.