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https://hdl.handle.net/2440/76227
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Type: | Journal article |
Title: | Activation of latent human GDF9 by a single residue change (Gly³⁹¹Arg) in the mature domain |
Other Titles: | Activation of latent human GDF9 by a single residue change (Gly(391)Arg) in the mature domain |
Author: | Simpson, C. Stanton, P. Walton, K. Chan, K. Ritter, L. Gilchrist, R. Harrison, C. |
Citation: | Endocrinology, 2012; 153(3):1301-1310 |
Publisher: | Endocrine Soc |
Issue Date: | 2012 |
ISSN: | 0013-7227 1945-7170 |
Statement of Responsibility: | Courtney M. Simpson, Peter G. Stanton, Kelly L. Walton, Karen L. Chan, Lesley J. Ritter, Robert B. Gilchrist, and Craig A. Harrison |
Abstract: | Growth differentiation factor 9 (GDF9) controls granulosa cell growth and differentiation during early ovarian folliculogenesis and regulates cumulus cell function and ovulation rate in the later stages of this process. Similar to other TGF-β superfamily ligands, GDF9 is secreted from the oocyte in a noncovalent complex with its prodomain. In this study, we show that prodomain interactions differentially regulate the activity of GDF9 across species, such that murine (m) GDF9 is secreted in an active form, whereas human (h) GDF9 is latent. To understand this distinction, we used site-directed mutagenesis to introduce nonconserved mGDF9 residues into the pro- and mature domains of hGDF9. Activity-based screens of the resultant mutants indicated that a single mature domain residue (Gly³⁹¹) confers latency to hGDF9. Gly³⁹¹ forms part of the type I receptor binding site on hGDF9, and this residue is present in all species except mouse, rat, hamster, galago, and possum, in which it is substituted with an arginine. In an adrenocortical cell luciferase assay, hGDF9 (Gly³⁹¹Arg) had similar activity to mGDF9 (EC₅₀ 55 ng/ml vs. 28 ng/ml, respectively), whereas wild-type hGDF9 was inactive. hGDF9 (Gly³⁹¹Arg) was also a potent stimulator of murine granulosa cell proliferation (EC₅₀ 52 ng/ml). An arginine at position 391 increases the affinity of GDF9 for its signaling receptors, enabling it to be secreted in an active form. This important species difference in the activation status of GDF9 may contribute to the variation observed in follicular development, ovulation rate, and fecundity between mammals. |
Keywords: | Adrenal Cortex Animals Humans Mice Arginine Mutagenesis, Site-Directed Cell Proliferation Amino Acid Sequence Protein Structure, Tertiary Sequence Homology, Amino Acid Ovulation Point Mutation Molecular Sequence Data Female Growth Differentiation Factor 9 HEK293 Cells |
Rights: | © 2012 by The Endocrine Society |
DOI: | 10.1210/en.2011-1632 |
Published version: | http://dx.doi.org/10.1210/en.2011-1632 |
Appears in Collections: | Aurora harvest 4 Obstetrics and Gynaecology publications |
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hdl_76227.pdf | Published version | 2.08 MB | Adobe PDF | View/Open |
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