An ELISA for SGP28/CRISP-3, a cysteine-rich secretory protein in human neutrophils, plasma, and exocrine secretions

Udby, Lene; Cowland, Jack B.; Johnsen, Anders H.; Sørensen, Ole E.; Borregaard, Niels; Kjeldsen, Lars

doi:10.1016/s0022-1759(02)00033-9

Cited by 83 publications

(80 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Human CRISP-3 was purified from granulocytes [10]. Specific antisera were raised in rabbits against recombinant C-terminally truncated human CRISP-3 and native human MSP [1,11]. Semen was collected from healthy volunteers and allowed to liquefy for 1 h at room temperature (RT), followed by centrifugation to remove spermatozoa and was subsequently stored at −20 °C until use.…”

Section: Methodsmentioning

confidence: 99%

“…We already knew that CRISP-3 is not covalently bound to other seminal plasma proteins, as separation of the proteins by SDS-PAGE followed by immunoblotting for CRISP-3 resulted in 2 bands corresponding to the expected sizes, also under non-reducing conditions [1]. The gel filtration experiments described above show that the MSP-CRISP-3 complex is stable and forms rapidly.…”

Section: Kinetics Of the Msp-crisp-3 Complex Formationmentioning

confidence: 97%

“…The concentration of CRISP-3 in seminal plasma was measured by enzyme-linked immunosorbent assay [1] and MSP with a modification of the competitive immunoassay for human MSP [11].…”

Section: Quantification Of Proteinsmentioning

confidence: 99%

“…Cysteine-rich secretory protein; β-Microseminoprotein; PSP94; Seminal plasma; Prostate cancer; Protein complex; SCP-domain; Mass spectrometry; Surface plasmon resonance Human cysteine-rich secretory protein 3 (CRISP-3; also known as SGP28) was first isolated from neutrophilic granulocytes, but it is also a constituent of blood plasma and exocrine secretions, such as saliva and seminal plasma [1]. It belongs to a family of CRISPs found in mammals and reptiles, which is characterized by 16 highly conserved cysteine residues of a total of 220-230 amino acids and 35-85% identity in the primary structure [2][3][4].…”

Section: Introductionmentioning

confidence: 99%

“…1A). CRISP-3 eluted as a double peak as we used a mixture of glycosylated and unglycosylated protein (shown in inset) with an apparent molecular mass of the glycosylated form estimated (from the calibration curve) to be 24 kDa, which is close to the expected monomeric size [1]. MSP eluted as a single peak corresponding to 21 kDa, suggesting that MSP either forms a homodimer or has an extended conformation under native conditions, since the theoretical mass of MSP is 10.7 kDa [7].…”

mentioning

confidence: 99%

See 4 more Smart Citations

β-Microseminoprotein binds CRISP-3 in human seminal plasma

Udby

Lundwall

Johnsen

et al. 2005

Biochemical and Biophysical Research Communications

Self Cite

View full text Add to dashboard Cite

Abstractβ -Microseminoprotein (MSP) and cysteine-rich secretory protein 3 (CRISP-3) are abundant constituents of human seminal plasma. Immunoprecipitation and gel filtration of seminal plasma proteins combined with examination of the proteins in their pure form showed that MSP and CRISP-3 form stable, non-covalent complexes. CRISP-3 binds MSP with very high affinity, as evidenced by surface plasmon resonance. Due to far higher abundance of MSP in prostatic fluid, it manifests large overcapacity for CRISP-3 binding. Structural similarity with an MSP-binding protein from blood plasma suggests that CRISP-3 binds MSP through its aminoterminal SCP-domain. KeywordsCysteine-rich secretory protein; β-Microseminoprotein; PSP94; Seminal plasma; Prostate cancer; Protein complex; SCP-domain; Mass spectrometry; Surface plasmon resonance Human cysteine-rich secretory protein 3 (CRISP-3; also known as SGP28) was first isolated from neutrophilic granulocytes, but it is also a constituent of blood plasma and exocrine secretions, such as saliva and seminal plasma [1]. It belongs to a family of CRISPs found in mammals and reptiles, which is characterized by 16 highly conserved cysteine residues of a total of 220-230 amino acids and 35-85% identity in the primary structure [2][3][4]. Recently, the crystal structure of a snake venom CRISP was elucidated [5]. This confirmed the previously proposed two-domain structure of CRISPs with a large N-terminal SCP domain (after sperm coating protein, an alternative name for murine CRISP-1) and a smaller, compact C-terminal domain, separated by a hinge-region [3,5].A novel protein with an N-terminal SCP domain has been identified in human blood plasma and is called PSP94-binding protein (PSPBP) as it binds to β -microseminoprotein (MSP), also known as prostate secretory protein of 94 amino acids (PSP94) [6]. MSP is one of the three predominant proteins secreted by the prostate gland, but it is also present in other body fluids, e.g., tracheobronchial fluid. In general, MSP-synthesis is associated with tissue producing [7,8].It is yet unknown whether MSP binds to PSPBP through the SCP domain or via the large C-terminal part of the molecule [6]. One way to answer this question is to investigate whether MSP binds other proteins with an SCP domain, and the abundance of both MSP and CRISP-3 in human seminal plasma prompted us to test the hypothesis that these proteins form high-affinity complexes. Materials and methods MaterialsHuman MSP was purified from seminal plasma with the method described for the purification of pig MSP [9]. Human CRISP-3 was purified from granulocytes [10]. Specific antisera were raised in rabbits against recombinant C-terminally truncated human CRISP-3 and native human MSP [1,11]. Semen was collected from healthy volunteers and allowed to liquefy for 1 h at room temperature (RT), followed by centrifugation to remove spermatozoa and was subsequently stored at −20 °C until use. Gel filtrationProteins were separated according to their molecular size by gel filtration on a Su...

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Kinetics Of the Msp-crisp-3 Complex Formationmentioning

confidence: 97%

“…The concentration of CRISP-3 in seminal plasma was measured by enzyme-linked immunosorbent assay [1] and MSP with a modification of the competitive immunoassay for human MSP [11].…”

Section: Quantification Of Proteinsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

β-Microseminoprotein binds CRISP-3 in human seminal plasma

Udby

Lundwall

Johnsen

et al. 2005

Biochemical and Biophysical Research Communications

Self Cite

View full text Add to dashboard Cite

show abstract

Low salivary dehydroepiandrosterone and androgen‐regulated cysteine‐rich secretory protein 3 levels in Sjögren's syndrome

Laine¹,

Porola²,

Udby³

et al. 2007

Arthritis & Rheumatism

Self Cite

View full text Add to dashboard Cite

Objective. Sjögren's syndrome (SS), an autoimmune disease of exocrine glands, typically starts at the time of adrenopause. We undertook this study to test the hypothesis that SS is characterized by an insufficient androgen effect at the target tissue level.Methods. We searched for androgen response elements (AREs) in the cysteine-rich secretory protein 3 (crisp-3) gene. Dehydroepiandrosterone (DHEA) responsiveness was experimentally studied using quantitative reverse transcriptase-polymerase chain reaction and immunofluorescence staining of human submandibular gland-derived acinar cells and labial salivary gland explants with or without DHEA. Finally, glandular and salivary CRISP-3 in healthy controls and SS patients was analyzed using immunohistochemistry, in situ hybridization, and enzyme-linked immunosorbent assay. Serum DHEA sulfate (DHEAS) and salivary DHEA levels were measured using a radioimmunometric method.Results. Literature analysis and a search for AREs in gene banks suggested androgen dependency of human CRISP-3, and this was verified by studies of human submandibular gland acinar cells cultured with or without DHEA, in which DHEA increased CRISP-3 messenger RNA (mRNA) levels (P ‫؍‬ 0.018). This finding was confirmed by the results of DHEA stimulation of labial salivary gland explants. Glandular CRISP-3 mRNA and protein labeling was weak and diffuse, coupled with low secretion in saliva (mean ؎ SEM 21.1 ؎ 2.7 g CRISP-3/15 minutes in SS patients versus 97.6 ؎ 12.0 g CRISP-3/15 minutes in healthy controls; P < 0.0001). Compared with healthy controls, SS patients had low serum levels of DHEAS (P ‫؍‬ 0.008) and also low salivary levels of DHEA (mean ؎ SEM 224 ؎ 33 pmoles versus 419 ؎ 98 pmoles; P ‫؍‬ 0.005).Conclusion. CRISP-3 pathology was seen in acini remote from lymphocyte foci and is apparently not secondary to local inflammation, but may represent some systemic effect in SS. Indeed, androgen deprivation in the salivary glands of SS patients is evidenced both by low salivary levels of DHEA and by low levels of DHEA-regulated CRISP-3. This may explain some of the characteristic features of SS.

show abstract