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Proteomic analysis of heparin-binding proteins from human seminal plasma: a step towards identification of molecular markers of male fertility

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Abstract

Glycosaminoglycans, especially heparin, are involved in various cell processes such as apoptosis, cell cycle control, platelet activation, capacitation, acrosome reaction and sperm decondensation. Heparin-binding proteins (HBPs) are essential constituents of human seminal fluid, which bind to sperm lipids containing the phosphorylcholine group and mediate the fertilization process. We utilized a proteomic set-up consisting of affinity chromatography, isoelectric focusing (IEF) coupled with matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI TOF/MS) for protein analysis of human HBPs. We resolved 70 different spots on two-dimensional (2-D) gel and subsequently identified these proteins. Forty different types of proteins were identified. Functional analysis revealed that 38% of the proteins belonged to the enzyme category, 20% were involved in RNA processing and transcription, 18% in structure and transport function, and 16% in cell recognition and signal transduction. We also identified 8% of proteins with unknown functions, although their expression in seminal fluid has been documented. Proteins of seminal fluid that bind heparin may be directly involved in sperm capacitation and acrosome reaction (AR), which are the two critical steps for fertilization. This information on HBPs would be useful for identifying potential biomarkers of fertility in the near future.

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Abbreviations

2-DE:

two-dimensional gel electrophoresis

ACN:

acetonitrile

ACTH:

adrenocortocotrophic hormone

AR:

acrosome reaction

DTT:

dithiothreitol

GAG:

glycosaminoglycan

HBP:

heparin-binding protein

IEF:

isoelectric focusing

IPG:

immobilized pH gradient

KRAB:

Kruppel-associated box

MALDI-TOF:

matrix-assisted laser desorption ionization time-of-flight

MS:

mass spectrometry

NCBI:

National Center for Biotechnology Information

PMF:

peptide mass fingerprinting

PSA:

prostate-specific antigen

TFA:

trifluoroacetic acid

ZFP:

zinc-finger protein

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Authors and Affiliations

  1. Department of Biophysics, All India Institute of Medical Sciences, New Delhi, 110 029, India

    Vijay Kumar, Md Imtaiyaz Hassan, Anil Kumar Tomar, Jaya Nautiyal, Tej P. Singh & Savita Yadav

  2. School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110 067, India

    Tara Kashav

  3. Department of Lab Medicine, All India Institute of Medical Sciences, New Delhi, 110 029, India

    Sarman Singh

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  1. Vijay Kumar
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  2. Md Imtaiyaz Hassan
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  3. Anil Kumar Tomar
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  4. Tara Kashav
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  5. Jaya Nautiyal
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  6. Sarman Singh
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  7. Tej P. Singh
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  8. Savita Yadav
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Correspondence to Savita Yadav.

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Kumar, V., Hassan, M.I., Tomar, A.K. et al. Proteomic analysis of heparin-binding proteins from human seminal plasma: a step towards identification of molecular markers of male fertility. J Biosci 34, 899–908 (2009). https://doi.org/10.1007/s12038-009-0104-5

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  • DOI: https://doi.org/10.1007/s12038-009-0104-5

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