Implantation Serine Proteinases heterodimerize and are critical in hatching and implantation

Sharma, Navneet; Liu, Shiying; Tang, Lin; Irwin, J.; Meng, Guoliang; Rancourt, Derrick E.

doi:10.1186/1471-213x-6-61

Cited by 40 publications

(47 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The supernatant was subjected to column chromatography using the Duo Flow FPLC system (BioRad) as per Sharma et al (2006). Protein purification involved sequential ion exchange (DEAE Sepharose) chromatography and two gel filtration (Superdex-75) steps.…”

Section: Protein Purificationmentioning

confidence: 99%

“…Thus, for purifying active recombinant ISP2, we turned to the use of conventional ion exchange chromatography followed by gel filtration, as was previously done for the purification of the native ISP complex from biological fluids (Sharma et al 2006) and recombinant ISP1 (Sharma et al 2011).…”

Section: Protein Purificationmentioning

confidence: 99%

“…The reaction conditions used were as per Sharma et al (2006) i.e., 25 mmol/L Tris.Cl (pH 8.0) buffer with 10 mmol/L EDTA reaction buffer, in a 0.5 mL total reaction volume including varying concentrations of the substrates and enzyme samples. The reactions were done at room temperature in a glass cuvette (1 cm path length) and scanned at wavelength of 405 nm (in case of p-nitroanilide conjugated substrate) and 540 nm (in case of methyl-naphthylamide substrates) using a spectrophotometer.…”

Section: Enzyme Assaymentioning

confidence: 99%

“…Subsequently it was discovered that both ISP1 and ISP2 are co-expressed as heterodimers (Sharma et al 2006). Inhibitors of implantation serine proteinase, as well as monoclonal antibodies raised against ISPs, are able to prevent murine embryo implantation (Sharma et al 2006;Huang et al 2004;Sun et al 2007) pointing to a key role for the ISPs in these critical processes of hatching and implantation.…”

Section: Introductionmentioning

confidence: 99%

“…Substrate specificity results using chromogenic p-nitroanilide conjugate substrates have demonstrated trypsin-like substrate specificity for the ISP1-ISP2 heterodimer (Sharma et al 2006). These results were confirmed with a phage display approach for establishing substrate preference (Sharma et al 2008).…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Implantation serine proteinase 2 is a monomeric enzyme with mixed serine proteolytic activity and can silence signalling via proteinase activated receptors

Sharma

Fahr

Renaux

et al. 2013

Biochem. Cell Biol.

Self Cite

View full text Add to dashboard Cite

Implantation serine proteinase 2 (ISP2), a S1 family serine proteinase, is known for its role in the critical processes of embryo hatching and implantation in the mouse uterus. Native implantation serine proteinases (ISPs) are co-expressed and co-exist as heterodimers in uterine and blastocyst tissues. The ISP1-ISP2 enzyme complex shows trypsin-like substrate specificity. In contrast, we found that ISP2, isolated as a 34 kDa monomer from a Pichia pastoris expression system, exhibited a mixed serine proteolytic substrate specificity, as determined by a phage display peptide cleavage approach and verified by the in vitro cleavage of synthetic peptides. Based upon the peptide sequence substrate selectivity, a database search identified many potential ISP2 targets of physiological relevance, including the proteinase activated receptor 2 (PAR2). The in vitro cleavage studies with PAR2-derived peptides confirmed the mixed substrate specificity of ISP2. Treatment of cell lines expressing proteinase-activated receptors (PARs) 1, 2, and 4 with ISP2 prevented receptor activation by either thrombin (PARs 1 and 4) or trypsin (PAR2). The disarming and silencing of PARs by ISP2 may play a role in successful embryo implantation.

show abstract

Section: Protein Purificationmentioning

confidence: 99%

Section: Protein Purificationmentioning

confidence: 99%

Section: Enzyme Assaymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Implantation serine proteinase 2 is a monomeric enzyme with mixed serine proteolytic activity and can silence signalling via proteinase activated receptors

Sharma

Fahr

Renaux

et al. 2013

Biochem. Cell Biol.

Self Cite

View full text Add to dashboard Cite

show abstract

Expression of rat uterine serine proteinases homologous to mouse implantation serine proteinase 2

et al. 2008

View full text Add to dashboard Cite

Implantation serine protease (ISP) was first identified in the uteri of pregnant mice. It is thought that ISP may have an important role in the initiation of implantation. However, the expression status and detailed functions of ISP remain unclear. In this study, the expression of ISP was investigated in the rat uterus. The analysis of two rat genes registered in GenBank, accession nos. XM_220240 and XM_577076, exhibited high identities to the mouse ISP2 genes, respectively at an mRNA level. We labeled the former as rISP2a and the latter as rISP2b. Using RT-PCR, we found that both genes were expressed in the uterus. Specifically, rISP2a mRNA was detected in the uterus throughout pregnancy, whereas rISP2b mRNA was only expressed in the uterus from day 5 of pregnancy until the end of gestation. Expression of both genes was observed specifically within the endometrial gland epithelium. Furthermore, rISP2a was also observed to be expressed in the fetus and placenta, whereas rISP2b expression was observed in the fetus but not in the placenta. An expressional signal of the rISP2a gene was observed in the spongiotrophoblasts, giant cells and decidual endometrium in the placenta. In the embryo, the ventral specific region was positive in rISP2a and rISP2b gene expression. These findings indicate the possibility that the presently examined genes with high identity to mouse ISP2 may play some role not only during the implantation phase, but also in the development of the placenta and embryo. J. Exp. Zool. (Mol. Dev. Evol.) 310B:642-649, 2008. r 2008 How to cite this article: Oozono S, Yamuchi N, Nishimura K, Matsumoto K, Watanabe R, Kubota K, Aramaki S, Sato F, Wood C, Soh T, Kizaki KI, Hattori MA. 2008. Expression of rat uterine serine proteinases homologous to mouse implantation serine proteinase 2. J. Exp. Zool. (Mol. Dev. Evol.) 310B:642-649.

show abstract

Characterization of secretory leukocyte protease inhibitor as an inhibitor of implantation serine proteinases

Sharma

Kaur

et al. 2007

Molecular Reproduction Devel

View full text Add to dashboard Cite

We have recently identified and characterized two implantation serine proteinase genes, ISP1 and ISP2, which give rise to a dimeric proteinase, ISP that facilitates embryo invasion during peri-implantation period. As many proteinases have cognate serpins that regulate their proteolytic activity, we have been investigating anti-tryptases, expressed during this window of implantation. Here, we report the differential expression of secretory leukocyte protease inhibitor (SLPI) in uterine endometrium around the implantation period. The co-localization of SLPI and ISP suggests the possibility that SLPI is an ISP serpin and that expression of SLPI may lead to a reduction in ISP activity. The expression of SLPI is down regulated during the window of embryo-uterine receptivity. Our results are consistent with a model suggesting that the drop in SLPI expression may help to refine the opening of the window of implantation, by allowing the proteolytic activity of embryo invasive serine proteinases such as the ISPs.

show abstract

Implantation Serine Proteinases heterodimerize and are critical in hatching and implantation

Abstract: Background: We have recently reported the expression of murine Implantation Serine Proteinase genes in pre-implantation embryos (ISP1) and uterus (ISP1 and ISP2). These proteinases belong to the S1 proteinase family and are similar to mast cell tryptases, which function as multimers.

Cited by 40 publications

References 28 publications

Implantation serine proteinase 2 is a monomeric enzyme with mixed serine proteolytic activity and can silence signalling via proteinase activated receptors

Implantation serine proteinase 2 is a monomeric enzyme with mixed serine proteolytic activity and can silence signalling via proteinase activated receptors

Expression of rat uterine serine proteinases homologous to mouse implantation serine proteinase 2

Characterization of secretory leukocyte protease inhibitor as an inhibitor of implantation serine proteinases

Contact Info

Product

Resources

About