Clin Exp Reprod Med.  2017 Jun;44(2):63-72. 10.5653/cerm.2017.44.2.63.

N-glycoproteomic analysis of human follicular fluid during natural and stimulated cycles in patients undergoing in vitro fertilization

• Lim HJ
• Seok AE
• Han J
• Lee J
• Lee S
• Kang HG
• Cha BH
• Yang Y

• KMID: 2405152
• DOI: http://doi.org/10.5653/cerm.2017.44.2.63

Abstract

OBJECTIVE
Hyperstimulation methods are broadly used for in vitro fertilization (IVF) in patients with infertility; however, the side effects associated with these therapies, such as ovarian hyperstimulation syndrome (OHSS), have not been well studied. N-glycoproteomes are subproteomes used for the remote sensing of ovarian stimulation in follicular growth. Glycoproteomic variation in human follicular fluid (hFF) has not been evaluated. In this study, we aimed to identify and quantify the glycoproteomes and N-glycoproteins (N-GPs) in natural and stimulated hFF using label-free nano-liquid chromatography/electrospray ionization-quad time-of-flight mass spectrometry.
METHODS
For profiling of the total proteome and glycoproteome, pooled protein samples from natural and stimulated hFF samples were selectively isolated using hydrazide chemistry to obtain the total proteomes and glycoproteomes. N-GPs were validated by the consensus sequence N-X-S/T (92.2% specificity for the N-glycomotif at p<0.05). All data were compared between natural versus hyperstimulated hFF samples.
RESULTS
We detected 41 and 44 N-GPs in the natural and stimulated hFF samples, respectively. Importantly, we identified 11 N-GPs with greater than two-fold upregulation in stimulated hFF samples compared to natural hFF samples. We also validated the novel N-GPs thyroxine-binding globulin, vitamin D-binding protein, and complement proteins C3 and C9.
CONCLUSION
We identified and classified N-GPs in hFF to improve our understanding of follicular physiology in patients requiring assisted reproduction. Our results provided important insights into the prevention of hyperstimulation side effects, such as OHSS.

Keyword

Fertilization in vitro; Human follicular fluid; Hyper stimulation; N-glycoprotein; Natural cycle; Proteomics

MeSH Terms

Chemistry
Complement System Proteins
Consensus Sequence
Female
Fertilization in Vitro*
Follicular Fluid*
Humans*
In Vitro Techniques*
Infertility
Mass Spectrometry
Ovarian Hyperstimulation Syndrome
Ovulation Induction
Physiology
Proteome
Proteomics
Reproduction
Sensitivity and Specificity
Thyroxine-Binding Globulin
Up-Regulation
Vitamin D-Binding Protein
Complement System Proteins
Proteome
Thyroxine-Binding Globulin
Vitamin D-Binding Protein