In-Depth Proteomic Analyses of Direct Expressed Prostatic Secretions

Drake, Richard; Elschenbroich, Sarah; Lopez-Perez, Orlay; Kim, Yunee; Ignatchenko, Vladimir; Ignatchenko, Alex; Nyalwidhe, Julius O.; Basu, Gaurav; Wilkins, Christopher E.; Gjurich, Breanne; Lance, Raymond S.; Semmes, O. John; Medin, Jeffrey A.; Kislinger, Thomas

doi:10.1021/pr1001498

Cited by 65 publications

(89 citation statements)

References 28 publications

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“…been detected more than 50 years ago. 15 Since then, more and more spots have been identified, and SP proteins corresponding to the same parent protein appear in multiple spots and bands, implying that there is a clear multiplicity of isoforms present, independently of the SP source (expressed prostate 49,50 ) or the bulk ejaculate. 51 Thousands of unique proteins have over time been identified, of which 25% were secretory.…”

Section: Proteomics Of the Ejaculatementioning

confidence: 99%

Seminal Plasma Proteins: What Role Do They Play?

Rodríguez‐Martínez

Kvist

Ernerudh

et al. 2011

American J Rep Immunol

326

273

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Citation Rodríguez‐Martínez H, Kvist U, Ernerudh J, Sanz L, Calvete JJ. Seminal Plasma Proteins: What Role Do They Play? Am J Reprod Immunol 2011; 66 (Suppl. 1): 11–22Problem Semen is a heterogenous and complex cell suspension in a protein‐rich fluid with different functions, some of them well known, others still obscure.Method of study This paper reviews, comparatively, our current knowledge on the growing field of proteomics of the SP and its relevance in relation to the in vivo situation, for the sake of reproductive biology, diagnostics and treatment.Results Ejaculated spermatozoa, primarily bathing in cauda epididymal fluid, are (in vitro) bulky, exposed to most, if not all, secretions from the accessory sexual glands. In vivo, however, not all spermatozoa are necessarily exposed to all secretions from these glands, because sperm cohorts are delivered in differential order and bathe in seminal plasma (SP) with different concentrations of constituents, including peptides and proteins. Proteins are relevant for sperm function and relate to sperm interactions with the various environments along the female genital tract towards the oocyte vestments. Specific peptides and proteins act as signals for the female immune system to modulate sperm rejection or tolerance, perhaps even influencing the relative intrinsic fertility of the male and/or couple by attaining a status of maternal tolerance towards embryo and placental development.Conclusions Proteins of the seminal plasma have an ample panorama of action, and some appear responsible for establishing fertility.

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Section: Proteomics Of the Ejaculatementioning

confidence: 99%

Seminal Plasma Proteins: What Role Do They Play?

Rodríguez‐Martínez

Kvist

Ernerudh

et al. 2011

American J Rep Immunol

326

273

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“…This method involves tryptic digestion of a protein mixture, and the resultant peptides are separated and identified by strong cation exchange (SCX) and reversed phase (RP) liquid chromatography coupled with mass spectrometry. MudPIT-based proteomics was applied to expressed prostatic secretions obtained from nine men with prostate cancer and resulted in the identification of 916 unique proteins [44].…”

Section: Technologies Involved In Proteomicsmentioning

confidence: 99%

Serum-based protein biomarkers for detection of lung cancer

2014

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Lung cancer is one of the most common cancers in terms of both incidence and mortality.The major reasons for the increasing number of deaths from lung cancer are late detection and lack of effective therapies. To improve our understanding of lung cancer biology, there is urgent need for blood-based, non-invasive molecular tests to assist in its detection in a cost-effective manner at an early stage when curative interventions are still possible. Recent advances in proteomic technology have provided extensive, high throughput analytical tools for identification, characterization and functional studies of proteomes. Changes in protein expression patterns in response to stimuli can serve as indicators or biomarkers of biological and pathological processes as well as physiological and pharmacological responses to drug treatment, thus aiding in early diagnosis and prognosis of disease. However, only a few biomarkers have been approved by the FDA to date for screening and diagnostic purposes. This review provides a brief overview of currently available proteomic techniques, their applications and limitations and the current state of knowledge about important serum biomarkers in lung cancer and their potential value as prognostic and diagnostic tools.

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“…Direct-EPS is a prostatic fluid that is collected from patients undergoing prostatectomy by massaging the organ and expelling 0.5-1 ml of the fluid just prior to surgical removal. It was chosen as our discovery fluid as it is expected to house prostate-secreted proteins at a higher concentration and purity, and we have developed a workflow for the in-depth proteomic analysis of this fluid (19). Following discovery proteomics in 16 clinically stratified direct-EPS samples, verification studies were performed using independent sample sets of direct-EPS.…”

mentioning

confidence: 99%

Identification of Differentially Expressed Proteins in Direct Expressed Prostatic Secretions of Men with Organ-confined Versus Extracapsular Prostate Cancer

Kim

Ignatchenko

Yao

et al. 2012

Molecular & Cellular Proteomics

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Current protocols for the screening of prostate cancer cannot accurately discriminate clinically indolent tumors from more aggressive ones. One reliable indicator of outcome has been the determination of organ-confined versus nonorgan-confined disease but even this determination is often only made following prostatectomy. This underscores the need to explore alternate avenues to enhance outcome prediction of prostate cancer patients. Fluids that are proximal to the prostate, such as expressed prostatic secretions (EPS), are attractive sources of potential prostate cancer biomarkers as these fluids likely bathe the tumor. Direct-EPS samples from 16 individuals with extracapsular (n ‫؍‬ 8) or organ-confined (n ‫؍‬ 8) prostate cancer were used as a discovery cohort, and were analyzed in duplicate by a nine-step MudPIT on a LTQ-Orbitrap XL mass spectrometer. A total of 624 unique proteins were identified by at least two unique peptides with a 0.2% false discovery rate. A semiquantitative spectral counting algorithm identified 133 significantly differentially expressed proteins in the discovery cohort. Integrative data mining prioritized 14 candidates, including two known prostate cancer biomarkers: prostate-specific antigen and prostatic acid phosphatase, which were significantly elevated in the direct-EPS from the organ-confined cancer group. These and five other candidates (SFN, MME, PARK7, TIMP1, and TGM4) were verified by Western blotting in an independent set of direct-EPS from patients with biochemically recurrent disease (n ‫؍‬ 5) versus patients with no evidence of recurrence upon follow-up (n ‫؍‬ 10). Lastly, we performed proof-of-concept SRM-MSbased relative quantification of the five candidates using unpurified heavy isotope-labeled synthetic peptides spiked into pools of EPS-urines from men with extracapsular and organ-confined prostate tumors. This study represents the first efforts to define the direct-EPS proteome from two major subclasses of prostate cancer using shotgun proteomics and verification in EPS-urine by SRM-MS. Molecular & Cellular

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In-Depth Proteomic Analyses of Direct Expressed Prostatic Secretions

Cited by 65 publications

References 28 publications

Seminal Plasma Proteins: What Role Do They Play?

Seminal Plasma Proteins: What Role Do They Play?

Serum-based protein biomarkers for detection of lung cancer

Identification of Differentially Expressed Proteins in Direct Expressed Prostatic Secretions of Men with Organ-confined Versus Extracapsular Prostate Cancer

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