Purification of a paracrine factor, P-Mod-S, produced by testicular peritubular cells that modulates Sertoli cell function

M K Skinner; P M Fetterolf; C T Anthony

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Purification of a paracrine factor, P-Mod-S, produced by testicular peritubular cells that modulates Sertoli cell function

Journal article

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Peer reviewed

Purification of a paracrine factor, P-Mod-S, produced by testicular peritubular cells that modulates Sertoli cell function

M K Skinner, P M Fetterolf and C T Anthony

The Journal of biological chemistry, Vol.263(6), pp.2884-2890

02/25/1988

Handle:

https://hdl.handle.net/2376/114824

PMID: 3343234

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Abstract

Testis - metabolism

Molecular Weight

Testis - cytology

Spermatogenesis

Electrophoresis, Polyacrylamide Gel

Amino Acids - analysis

Fluorometry

Androgen-Binding Protein - metabolism

Dose-Response Relationship, Drug

Sertoli Cells - physiology

Animals

Testicular Hormones - isolation & purification

Transferrin - metabolism

Reproduction

A testicular paracrine factor, P-Mod-S, was purified from conditioned medium obtained from serum-free cultures of peritubular cells. Stimulation of testicular transferrin production by cultured Sertoli cells was utilized as a bio-assay for P-Mod-S. A bioactive protein with an apparent molecular weight of 50,000 under physiological conditions was isolated by high pressure size exclusion chromatography. P-Mod-S was found to have an affinity for heparin and bound to a heparin affinity column. Two forms of P-Mod-S were purified with reverse-phase chromatography. The less hydrophobic form was referred to as P-Mod-S (A) and is a 56,000 molecular weight protein. The more hydrophobic form was referred to as P-Mod-S (B) and is a 59,000 molecular weight protein. Purification of P-Mod-S (A) and P-Mod-S (B) from peritubular cell-radiolabeled secreted proteins revealed that both proteins contain radioactivity. This result demonstrates active synthesis and secretion of P-Mod-S by peritubular cells. Although the amino acid composition of the two proteins indicates distinct differences in the content of several amino acids, the relationship of P-Mod-S (A) and P-Mod-S (B) is unknown at present. A greater than 1000-fold increase in the specific activity of P-Mod-S was achieved with the purification procedure utilized. P-Mod-S can account for essentially all the bioactivity present in crude peritubular cell-secreted protein preparations. The effects of the two forms of P-Mod-S on both transferrin and androgen-binding protein production by Sertoli cells was examined. Purified forms of P-Mod-S were found to have a greater effect on Sertoli cell function than any individual regulatory agent previously known to influence the cell, including follicle-stimulating hormone. The significance of peritubular cell-Sertoli cell interactions mediated via P-Mod-S to spermatogenesis and testicular function is discussed, as well as insight provided into general mesenchymal-epithelial cell interactions.

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Title: Purification of a paracrine factor, P-Mod-S, produced by testicular peritubular cells that modulates Sertoli cell function
Creators: M K Skinner - Department of Pharmacology, Vanderbilt University, School of Medicine, Nashville, Tennessee 37232
P M Fetterolf
C T Anthony
Publication Details: The Journal of biological chemistry, Vol.263(6), pp.2884-2890
Academic Unit: Biological Sciences, School of
Publisher: United States
Grant note: T32 HD-07043 / NICHD NIH HHS HD-20583 / NICHD NIH HHS
Identifiers: 99900548224101842
Language: English
Resource Type: Journal article