BACKGROUND: Gamma-glutamyltransferase (GGT) is a well-known laboratory biomarker. In spite of high concentration and the possible biomedical importance of estimating GGT in human seminal plasma (hSP), it has not been widely explored in reproductive physiology. This study aimed to complement existing data on its diversity, previously obtained on seminal extracellular vesicles, by analyzing matched soluble fraction of hSP. The GGT-associated patterns of selected glycoproteins were analyzed in order to establish an adjunct referent parameter for differentiation between known high molecular mass forms of GGT. Getting insight into distinct GGT-associated glycoprotein patterns should contribute to define them together as possible multimarkers. METHODS: GGT forms in soluble, membrane-free-fraction isolated form hSP of normozoospermic men were analyzed using gel filtration and lectin blotting using WGA (wheat germ agglutinin) and Con A (concanavalin A). RESULTS: Widely distributed GGT (with two to three partially resolved peaks), which may correspond to high molecular mass aggregates, were detected. GGT-associated patterns of selected glycoproteins (at position of big, medium, and small-GGT) all comprised high molecular mass WGA-reactive smears, but differed in the presence of Con A-reactive glycans, as well as mucin-associated antigens CA19-9 and CA125. CONCLUSIONS: GGT contributes to several molecular patterns that differ between the soluble and extracellular vesicle fractions of hSP. Their glycobiochemical heterogeneity is due to difference in the presence of distinct sialylated and mannosylated glycans. Moreover, GGT-associated glycoprotein patterns differentiate between high molecular mass forms of GGT in the soluble fraction of hSP. They hold promise as possible targets for increasing biomarker potential of GGT.

Department for Immunochemistry and Glycobiology Institute for the Application of Nuclear Energy INEP University of Belgrade Belgrade Serbia

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Heisterkamp N, Groffen J, Warburton D, Sneddon T. The human gamma-glutamyltransferase gene family. Hum Genet 2008;123:321-32. DOI

Nemesanszky E, Lott JA. Gamma-glutamyltransferase and its isoenzymes: Progress and problems. Clin Chem 1985:797-803. DOI

Balakrishna S, Prabhune AA. Gamma-glutamyl transferases: A structural, mechanistic and physiological perspective. Front Biol (Beijing) 2014;9:51-65. DOI

Castellano I, Merlino A. γ-Glutamyltranspeptidases: sequence, structure, biochemical properties, and biotechnological applications. Cell Mol Life Sci 2012;69:3381-94. DOI

Selvaraj P, Rolston DDK, Balasubramanian KA. Separation of hydrophobic and hydrophilic forms of γ-glutamyltransferase from human serum by hydrophobic chromatography on phenyl-Sepharose CL-4B: studies on normal sera and sera of patients with liver disease. Clin Chim Acta 1984;138:141-9. DOI

West MB, Segu ZM, Feasley CL, Kang P, Klouckova I, Li C, Novotny M V., West CM, Mechref Y, Hanigan MH. Analysis of site-specific glycosylation of renal and hepatic γ-glutamyl transpeptidase from normal human tissue. J Biol Chem 2010;285:29511-24. DOI

Tate SS, Khadse V. Renal γ-glutamyl transpeptidases: Influence of glycosylation on the electrophoretic behavior and molecular weights of their subunits. Biochem Biophys Res Commun 1986;141:1189-94. DOI

Hughey RP, Curthoys NP. Comparison of the size and physical properties of γ glutamyltranspeptidase purified from rat kidney following solubilization with papain or with triton X 100. J Biol Chem 1976;251:7863-70. DOI

David Castle J, Cameron RS, Patterson PL, Ma AK. Identification of high molecular weight antigens structurally related to gamma-glutamyl transferase in epithelial tissues. J Membr Biol 1985;87:13-26. DOI

Corti A, Franzini M, Cianchetti S, Bergamini G, Lorenzini E, Melotti P, Paolicchi A, Paggiaro P, Pompella A. Contribution by polymorphonucleate granulocytes to elevated gamma-glutamyltransferase in cystic fibrosis sputum. PLoS One 2012;7:e34772. DOI

Ingbar DH, Hepler K, Dowin R, Jacobsen E, Dunitz JM, Nici L, Jamieson JD. γ-Glutamyl transpeptidase is a polarized alveolar epithelial membrane protein. Am J Physiol - Lung Cell Mol Physiol 1995;269. DOI

Whitfield JB. Gamma glutamyl transferase. Crit Rev Clin Lab Sci 2001;38:263-355. DOI

Corti A, Paolicchi A, Franzini M, Dominici S, Casini AF, Pompella A. The S-thiolating activity of membrane γ-glutamyltransferase: Formation of cysteinyl-glycine mixed disulfides with cellular proteins and in the cell microenvironment. Antioxidants Redox Signal 2005;7:911-8. DOI

Turgut O, Tandogan I. Gamma-glutamyltransferase to determine cardiovascular risk: shifting the paradigm forward. J Atheroscler Thromb 2011;18:177-81. DOI

Franzini M, Bramanti E, Ottaviano V, Ghiri E, Scatena F, Barsacchi R, Pompella A, Donato L, Emdin M, Paolicchi A. A high performance gel filtration chromatography method for γ-glutamyltransferase fraction analysis. Anal Biochem 2008;374:1-6. DOI

Fornaciari I, Fierabracci V, Corti A, Elawadi HA, Lorenzini E, Emdin M, Paolicchi A, Franzini M. Gamma-glutamyltransferase fractions in human plasma and bile: Characteristic and biogenesis. PLoS One 2014;9:e88532. DOI

Remaley A, Wilding P. Macroenzymes: biochemical characterization, clinical significance, and laboratory detection. Clin Chem 1989;35:2261-70. DOI

Foddis R, Franzini M, Bonotti A, Marino R, Silvestri R, Fallahi P, Chiappino D, Emdin M, Paolicchi A, Cristaudo A. Big and Free Fractions of Gamma-Glutamyltransferase: New Diagnostic Biomarkers for Malignant Mesothelioma? Diagnostics 2022;12:311. DOI

Franzini M, Fornaciari I, Rong J, Larson MG, Passino C, Emdin M, Paolicchi A, Vasan RS. Correlates and reference limits of plasma gamma-glutamyltransferase fractions from the Framingham Heart Study. Clin Chim Acta 2013;417:19-25. DOI

Franzini M, Fornaciari I, Vico T, Moncini M, Cellesi V, Meini M, Emdin M, Paolicchi A. High-sensitivity gamma-glutamyltransferase fraction pattern in alcohol addicts and abstainers. Drug Alcohol Depend 2013;127:239-42. DOI

Verhoeven G, Steeno O. Evidence for the prostatic origin of gamma-glutamyltransferase acivity in human semen. Andrologia 1979;1:163-9.

Verze P, Cai T, Lorenzetti S. The role of the prostate in male fertility, health and disease. Nat Rev Urol 2016;13:379-86. DOI

Chen F, Lu JC, Xu HR, Huang YF, Hu YA SY. Evaluation of the determination of seminal ACP and gamma-GT activities and correlation between seminal ACP or gamma-GT activity and semen parameters. Zhonghua Nan Ke Xue 2006;12:879-82.

Aafjes J, Oort H, Schenck P. Gamma-glutamyltranspeptidase and β-glucuronidase in human ejaculates. relation with abnormal liquefaction. Andrologia 1983;15:661-5. DOI

Djordjevic D, Lalic N, Vukovic I, Nale D. Sperm quality and seminal biochemical parameters in infertile men with and without leukocytospermia. Andrology 2018;7:2167-250.

Ehala-Aleksejev K, Punab M. Serum hepatic enzyme activity in relation to semen quality and serum reproductive hormone levels among Estonian fertile Men. Andrology 2016;4:152-9. DOI

Feng R, Lu J, Zhang H, Lü N. A pilot comparative study of 26 biochemical markers in seminal plasma and serum in infertile men. Biomed Res Int 2015;2015:805328. doi: 10.1155/2015/805328 PubMed DOI

Janković T, Goč S, Mitić N, Danilović Luković J, Janković M. Membrane-associated gamma-glutamyl transferase and alkaline phosphatase in the context of concanavalin A- and wheat germ agglutinin-reactive glycans mark seminal prostasome populations from normozoospermic and oligozoospermic men. Ups J Med Sci 2020:125(1):10-8. DOI

Janković T, Luković JD, Miler I, Mitić N, Hajduković L, Janković M. Assembly of tetraspanins, galectin-3, and distinct N-glycans defines the solubilization signature of seminal prostasomes from normozoospermic and oligozoospermic men. Ups J Med Sci 2021 Sep 3;126. doi: 10.48101/ujms.v126.7673 PubMed DOI

Janković M. Glycans as biomarkers: status and perspectives. J Med Biochem 2011;30:213-23. DOI

Carlsson L, Nilsson O, Larsson A, Stridsberg M, Sahlén G, Ronquist G. Characteristics of human prostasomes isolated from three different sources. Prostate 2003;54:322-30. DOI

Davidson DF, Watson DJM. Macroenzyme detection by polyethylene glycol precipitation. Ann Clin Biochem 2003;40:514-20. DOI

Zhang X, Borg EGF, Liaci AM, Vos HR, Stoorvogel W. A novel three step protocol to isolate extracellular vesicles from plasma or cell culture medium with both high yield and purity. J Extracell Vesicles 2020;9(1):1791450. doi: 10.1080/20013078.2020.1791450 PubMed DOI

Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227:680-5. DOI

Huseby NE. Multiple forms of γ-glutamyltransferase in normal human liver, bile and serum. Biochim Biophys Acta (BBA). Enzymology 1978;522:354-62. DOI

Arai K, Yoshida KI, Komoda T, Sakagishi Y. Comparative studies on the properties of purified gamma-glutamyl transferase from human reproductive system and the kidney. Clin Biochem 1990;23:105-12. DOI

Kimura Y, Kido A, Oya M. Phenotyping of Gamma-Glutamyl Transferase in Human Semen, the Prostate and Seminal Stains. Yamanashi Med J 1994;9:97-103.

Abe S, Kochi H, Hiraiwa K. Purification and immunological characterization of a new form of γ-glutamyltransferase of human semen. Biochim Biophys Acta (BBA)-Protein Struct Mol Enzymol 1991;1077:259-64. DOI

Franzini M, Ottaviano V, Fierabracci V, Bramanti E, Zyw L, Barsacchi R, Scatena F, Boni C, Mammini C, Passino C, Pompella A, Emdin M, A P. Fractions of plasma gamma-glutamyltransferase in healthy individuals: reference values. Clin Chim Acta 2008;395:188-9. DOI

Bromw J, Raulf M, Stuning M, Spurt B, Creat A, Bremm KD, Lehrstuhl WK, Konig W. Subcellular localization of enzymes involved in leukotriene formation within human polymorphonuclear granulocytes. Immunology 1984;51:571.

Raulf M, Stüning M, Köni W. Metabolism of leukotrienes by L-gamma-glutamyl-transpeptidase and dipeptidase from human polymorphonuclear granulocytes. ImmunologyImmunology 1985;55:135.

Cocucci E, Racchetti G, Meldolesi J. Shedding microvesicles: artefacts no more. Trends Cell Biol 2009;19:43-51. DOI

Zhou B, Xu K, Zheng X, Chen T, Wang J, Song Y, Shao Y, Zheng S. Application of exosomes as liquid biopsy in clinical diagnosis. Signal Transduct Target Ther 2020;5:144. DOI

Valencia K, Montuenga LM. Exosomes in Liquid Biopsy: The Nanometric World in the Pursuit of Precision Oncology. Cancers (Basel) 2021;13:2147. DOI

Radicioni G, Cao R, Carpenter J, Ford AA, Wang TT, Li Y, Kesimer M. The innate immune properties of airway mucosal surfaces are regulated by dynamic interactions between mucins and interacting proteins: the mucin interactome. Mucosal Immunol 2016;9:1442-54. DOI

Zhang H, Forman H, Choi J. γ-Glutamyl transpeptidase in glutathione biosynthesis. Methods Enzymol 2005;401:468-83. DOI

Lyu S, Wang J, Huang M, Wang H, Zhou L, He Q, Lang R. CA19-9 Level to Serum γ-Glutamyltransferase as a Potential Prognostic Biomarker in Patients with Pancreatic Head Carcinoma. Cancer Menag Res 2021;13:4887. DOI

Wang F, Lyu S, Cao D, Ren Z, Zhao X, Li L, He Q. Correlation between preoperative CA19-9/GGT ratio and postoperative long-term survival of distal cholangiocarcinoma. Chinese J Hepatobiliary Surg 2021:762-6.