TY  - CONF
AU  - Danilović Luković, Jelena
AU  - Janković, Tamara
AU  - Goč, Sanja
AU  - Janjić, Filip
AU  - Mitic, Ninoslav
PY  - 2023
UR  - http://rimsi.imsi.bg.ac.rs/handle/123456789/2732
AB  - Glycans, complex carbohydrates bound to lipids and proteins, play important roles in biological processes.
Prostasomes, extracellular vesicles from prostate epithelial cells, have a glycan composition influenced by
their cellular origin and by the composition of the seminal plasma into which they are secreted. We
hypothesized that the membrane-associated glycans could be used as selective targets for separation of
prostasomes and as relevant parameter for distinction of their populations.
Prostasomes from seminal plasma of normozoospermic and oligozoospermic men were separated by
ion-exchange- and lectin-affinity chromatography using concanavalin A (ConA) lectin, specific for
mannosylated structures, and wheat germ agglutinin (WGA), specific for sialic acid. The presence of
tetraspanins (CD63, CD9 and CD81), galectin-3 (gal-3) and gamma-glutamyl transferase (GGT) as known
vesicle markers were monitored in association with distinct glycans. Their distribution was also analysed upon
treatment of prostasomes with non-ionic detergent.
Membrane-associated GGT in the context of Con A- and WGA-reactive glycans mark prostasome
populations from normozoospermic and oligozoospermic men. The assembly of tetraspanins, gal-3, and
distinct N-glycans defines the solubilisation signature of prostasomes. WGA-reactive glycoproteins
co-localize with CD9 and gal-3 in detergent-resistant domains, whereas ConA-reactive glycoproteins were
distributed in detergent-sensitive domains along with CD63 and GGT. Subtle differences in the
composition/presentation of examined molecules made difference among vesicles sharing the same physical properties in each group as well as between them.
The results obtained suggest the potential of glyco-parameters as reference markers for EVs populations.
PB  - Medical University Graz, Austria
C3  - Joint Meeting of ASEV, HSEV, SiNEV and SrbEV, Small New World 2.0, Graz, Austria
T1  - Surface-Associated Glycans as a Possible Distinct Factor for Establishing the Molecular Properties of Prostasomes
EP  - 52
SP  - 52
UR  - https://hdl.handle.net/21.15107/rcub_rimsi_2732
ER  - 

@conference{
author = "Danilović Luković, Jelena and Janković, Tamara and Goč, Sanja and Janjić, Filip and Mitic, Ninoslav",
year = "2023",
abstract = "Glycans, complex carbohydrates bound to lipids and proteins, play important roles in biological processes.
Prostasomes, extracellular vesicles from prostate epithelial cells, have a glycan composition influenced by
their cellular origin and by the composition of the seminal plasma into which they are secreted. We
hypothesized that the membrane-associated glycans could be used as selective targets for separation of
prostasomes and as relevant parameter for distinction of their populations.
Prostasomes from seminal plasma of normozoospermic and oligozoospermic men were separated by
ion-exchange- and lectin-affinity chromatography using concanavalin A (ConA) lectin, specific for
mannosylated structures, and wheat germ agglutinin (WGA), specific for sialic acid. The presence of
tetraspanins (CD63, CD9 and CD81), galectin-3 (gal-3) and gamma-glutamyl transferase (GGT) as known
vesicle markers were monitored in association with distinct glycans. Their distribution was also analysed upon
treatment of prostasomes with non-ionic detergent.
Membrane-associated GGT in the context of Con A- and WGA-reactive glycans mark prostasome
populations from normozoospermic and oligozoospermic men. The assembly of tetraspanins, gal-3, and
distinct N-glycans defines the solubilisation signature of prostasomes. WGA-reactive glycoproteins
co-localize with CD9 and gal-3 in detergent-resistant domains, whereas ConA-reactive glycoproteins were
distributed in detergent-sensitive domains along with CD63 and GGT. Subtle differences in the
composition/presentation of examined molecules made difference among vesicles sharing the same physical properties in each group as well as between them.
The results obtained suggest the potential of glyco-parameters as reference markers for EVs populations.",
publisher = "Medical University Graz, Austria",
journal = "Joint Meeting of ASEV, HSEV, SiNEV and SrbEV, Small New World 2.0, Graz, Austria",
title = "Surface-Associated Glycans as a Possible Distinct Factor for Establishing the Molecular Properties of Prostasomes",
pages = "52-52",
url = "https://hdl.handle.net/21.15107/rcub_rimsi_2732"
}

Danilović Luković, J., Janković, T., Goč, S., Janjić, F.,& Mitic, N.. (2023). Surface-Associated Glycans as a Possible Distinct Factor for Establishing the Molecular Properties of Prostasomes. in Joint Meeting of ASEV, HSEV, SiNEV and SrbEV, Small New World 2.0, Graz, Austria
Medical University Graz, Austria., 52-52.
https://hdl.handle.net/21.15107/rcub_rimsi_2732

Danilović Luković J, Janković T, Goč S, Janjić F, Mitic N. Surface-Associated Glycans as a Possible Distinct Factor for Establishing the Molecular Properties of Prostasomes. in Joint Meeting of ASEV, HSEV, SiNEV and SrbEV, Small New World 2.0, Graz, Austria. 2023;:52-52.
https://hdl.handle.net/21.15107/rcub_rimsi_2732 .

Danilović Luković, Jelena, Janković, Tamara, Goč, Sanja, Janjić, Filip, Mitic, Ninoslav, "Surface-Associated Glycans as a Possible Distinct Factor for Establishing the Molecular Properties of Prostasomes" in Joint Meeting of ASEV, HSEV, SiNEV and SrbEV, Small New World 2.0, Graz, Austria (2023):52-52,
https://hdl.handle.net/21.15107/rcub_rimsi_2732 .

TY  - JOUR
AU  - Janković, Tamara
AU  - Danilović Luković, Jelena
AU  - Goč, Sanja
AU  - Mitić, Ninoslav
AU  - Hajduković, Ljiljana
AU  - Janković, Miroslava
PY  - 2023
UR  - http://rimsi.imsi.bg.ac.rs/handle/123456789/2497
AB  - 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.
PB  - Faculty of Medicine and Dentistry Palacký University Olomouc, Czech Republic
T2  - Biomedical papers of the Medical Faculty of the University Palacky, Olomouc, Czechoslovakia
T1  - Gamma-glutamyltransferase-associated glycoprotein patterns in human seminal plasma of normozoospermic men: a new aspect of biomarker heterogeneity
DO  - 10.5507/bp.2023.031
ER  - 

@article{
author = "Janković, Tamara and Danilović Luković, Jelena and Goč, Sanja and Mitić, Ninoslav and Hajduković, Ljiljana and Janković, Miroslava",
year = "2023",
abstract = "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.",
publisher = "Faculty of Medicine and Dentistry Palacký University Olomouc, Czech Republic",
journal = "Biomedical papers of the Medical Faculty of the University Palacky, Olomouc, Czechoslovakia",
title = "Gamma-glutamyltransferase-associated glycoprotein patterns in human seminal plasma of normozoospermic men: a new aspect of biomarker heterogeneity",
doi = "10.5507/bp.2023.031"
}

Janković, T., Danilović Luković, J., Goč, S., Mitić, N., Hajduković, L.,& Janković, M.. (2023). Gamma-glutamyltransferase-associated glycoprotein patterns in human seminal plasma of normozoospermic men: a new aspect of biomarker heterogeneity. in Biomedical papers of the Medical Faculty of the University Palacky, Olomouc, Czechoslovakia
Faculty of Medicine and Dentistry Palacký University Olomouc, Czech Republic..
https://doi.org/10.5507/bp.2023.031

Janković T, Danilović Luković J, Goč S, Mitić N, Hajduković L, Janković M. Gamma-glutamyltransferase-associated glycoprotein patterns in human seminal plasma of normozoospermic men: a new aspect of biomarker heterogeneity. in Biomedical papers of the Medical Faculty of the University Palacky, Olomouc, Czechoslovakia. 2023;.
doi:10.5507/bp.2023.031 .

Janković, Tamara, Danilović Luković, Jelena, Goč, Sanja, Mitić, Ninoslav, Hajduković, Ljiljana, Janković, Miroslava, "Gamma-glutamyltransferase-associated glycoprotein patterns in human seminal plasma of normozoospermic men: a new aspect of biomarker heterogeneity" in Biomedical papers of the Medical Faculty of the University Palacky, Olomouc, Czechoslovakia (2023),
https://doi.org/10.5507/bp.2023.031 . .

TY  - JOUR
AU  - Danilović Luković, Jelena
AU  - Goč, Sanja
AU  - Mitic, Ninoslav
AU  - Janković, Tamara
PY  - 2022
UR  - http://rimsi.imsi.bg.ac.rs/handle/123456789/2476
AB  - The carbohydrate portion of proteins and lipids mediates a variety of biological processes. Revealing their
underlying principles is a challenging task that could contribute to a better understanding of many patho/physiological
conditions. On the other hand, the interest in extracellular vesicles (EVs) has increased in recent years due to their
involvement in intercellular communication leading to an array of functional and structural changes in recipient cells.
Their characterization uncovered an exceptional diversity in size, morphology, as well as in membrane and cargo content.
Monitoring/analysis of surface glycosylation of EVs originating from the prostate, termed prostasomes, revealed
their substantial contribution to the complexity of seminal plasma (SP) glycome. Heterogeneity of surface glycans confirm
the existence of several prostasome subpopulations. Presentation of surface glycans on prostasomal membrane is
strongly affected by co-localized membrane-associated glycoproteins and tetraspanins. They appear to be organized in
established/regular distribution patterns on membrane domains. Surface glycans are a component of EVs membrane
that affects its functionality and potentially a distinction marker of prostasome subpopulations. Further understanding
of the complex composition of glycans on EVs might explain the relation of their structure with functional alterations
in distinct patho/physiological conditions.
PB  - Faculty of Sciences, Department of Biology, University of Novi Sad, Serbia
T2  - Biologia Serbica
T1  - Extracellular vesicles in a maze of glycomic complexity
EP  - 32
IS  - 1
SP  - 25
VL  - 44
DO  - 10.5281/zenodo.7075050
ER  - 

@article{
author = "Danilović Luković, Jelena and Goč, Sanja and Mitic, Ninoslav and Janković, Tamara",
year = "2022",
abstract = "The carbohydrate portion of proteins and lipids mediates a variety of biological processes. Revealing their
underlying principles is a challenging task that could contribute to a better understanding of many patho/physiological
conditions. On the other hand, the interest in extracellular vesicles (EVs) has increased in recent years due to their
involvement in intercellular communication leading to an array of functional and structural changes in recipient cells.
Their characterization uncovered an exceptional diversity in size, morphology, as well as in membrane and cargo content.
Monitoring/analysis of surface glycosylation of EVs originating from the prostate, termed prostasomes, revealed
their substantial contribution to the complexity of seminal plasma (SP) glycome. Heterogeneity of surface glycans confirm
the existence of several prostasome subpopulations. Presentation of surface glycans on prostasomal membrane is
strongly affected by co-localized membrane-associated glycoproteins and tetraspanins. They appear to be organized in
established/regular distribution patterns on membrane domains. Surface glycans are a component of EVs membrane
that affects its functionality and potentially a distinction marker of prostasome subpopulations. Further understanding
of the complex composition of glycans on EVs might explain the relation of their structure with functional alterations
in distinct patho/physiological conditions.",
publisher = "Faculty of Sciences, Department of Biology, University of Novi Sad, Serbia",
journal = "Biologia Serbica",
title = "Extracellular vesicles in a maze of glycomic complexity",
pages = "32-25",
number = "1",
volume = "44",
doi = "10.5281/zenodo.7075050"
}

Danilović Luković, J., Goč, S., Mitic, N.,& Janković, T.. (2022). Extracellular vesicles in a maze of glycomic complexity. in Biologia Serbica
Faculty of Sciences, Department of Biology, University of Novi Sad, Serbia., 44(1), 25-32.
https://doi.org/10.5281/zenodo.7075050

Danilović Luković J, Goč S, Mitic N, Janković T. Extracellular vesicles in a maze of glycomic complexity. in Biologia Serbica. 2022;44(1):25-32.
doi:10.5281/zenodo.7075050 .

Danilović Luković, Jelena, Goč, Sanja, Mitic, Ninoslav, Janković, Tamara, "Extracellular vesicles in a maze of glycomic complexity" in Biologia Serbica, 44, no. 1 (2022):25-32,
https://doi.org/10.5281/zenodo.7075050 . .

TY  - JOUR
AU  - Janković, Tamara
AU  - Danilović Luković, Jelena
AU  - Miler, Irena
AU  - Mitic, Ninoslav
AU  - Hajduković, Ljiljana
AU  - Janković, Miroslava
PY  - 2021
UR  - http://rimsi.imsi.bg.ac.rs/handle/123456789/2464
AB  - Background: Prostasomes, extracellular vesicles (EVs) abundantly present in seminal plasma, express distinct
tetraspanins (TS) and galectin-3 (gal-3), which are supposed to shape their surface by an assembly of
different molecular complexes. In this study, detergent-sensitivity patterns of membrane-associated prostasomal
proteins were determined aiming at the solubilization signature as an intrinsic multimolecular
marker and a new parameter suitable as a reference for the comparison of EVs populations in health and
disease.
Methods: Prostasomes were disrupted by Triton X-100 and analyzed by gel filtration under conditions that
maintained complete solubilization. Redistribution of TS (CD63, CD9, and CD81), gal-3, gamma-glutamyltransferase
(GGT), and distinct N-glycans was monitored using solid-phase lectin-binding assays, transmission
electron microscopy, electrophoresis, and lectin blot.
Results: Comparative data on prostasomes under normal physiology and conditions of low sperm count
revealed similarity regarding the redistribution of distinct N-glycans and GGT, all presumed to be mainly
part of the vesicle coat. In contrast to this, a greater difference was found in the redistribution of integral
membrane proteins, exemplified by TS and gal-3. Accordingly, they were grouped into two molecular patterns
mainly consisting of overlapped CD9/gal-3/wheat germ agglutinin-reactive glycoproteins and CD63/
GGT/concanavalin A-reactive glycoproteins.
Conclusions: Solubilization signature can be considered as an all-inclusive distinction factor regarding the
surface properties of a particular vesicle since it reflects the status of the parent cell and the extracellular
environment, both of which contribute to the composition of spatial membrane arrangements.
PB  - Upsala Medical Society
T2  - Upsala journal of medical sciences
T1  - Assembly of tetraspanins, galectin-3, and distinct N-glycans defines the solubilization signature of seminal prostasomes from normozoospermic and oligozoospermic men
VL  - 126
DO  - 10.48101/ujms.v126.7673
ER  - 

@article{
author = "Janković, Tamara and Danilović Luković, Jelena and Miler, Irena and Mitic, Ninoslav and Hajduković, Ljiljana and Janković, Miroslava",
year = "2021",
abstract = "Background: Prostasomes, extracellular vesicles (EVs) abundantly present in seminal plasma, express distinct
tetraspanins (TS) and galectin-3 (gal-3), which are supposed to shape their surface by an assembly of
different molecular complexes. In this study, detergent-sensitivity patterns of membrane-associated prostasomal
proteins were determined aiming at the solubilization signature as an intrinsic multimolecular
marker and a new parameter suitable as a reference for the comparison of EVs populations in health and
disease.
Methods: Prostasomes were disrupted by Triton X-100 and analyzed by gel filtration under conditions that
maintained complete solubilization. Redistribution of TS (CD63, CD9, and CD81), gal-3, gamma-glutamyltransferase
(GGT), and distinct N-glycans was monitored using solid-phase lectin-binding assays, transmission
electron microscopy, electrophoresis, and lectin blot.
Results: Comparative data on prostasomes under normal physiology and conditions of low sperm count
revealed similarity regarding the redistribution of distinct N-glycans and GGT, all presumed to be mainly
part of the vesicle coat. In contrast to this, a greater difference was found in the redistribution of integral
membrane proteins, exemplified by TS and gal-3. Accordingly, they were grouped into two molecular patterns
mainly consisting of overlapped CD9/gal-3/wheat germ agglutinin-reactive glycoproteins and CD63/
GGT/concanavalin A-reactive glycoproteins.
Conclusions: Solubilization signature can be considered as an all-inclusive distinction factor regarding the
surface properties of a particular vesicle since it reflects the status of the parent cell and the extracellular
environment, both of which contribute to the composition of spatial membrane arrangements.",
publisher = "Upsala Medical Society",
journal = "Upsala journal of medical sciences",
title = "Assembly of tetraspanins, galectin-3, and distinct N-glycans defines the solubilization signature of seminal prostasomes from normozoospermic and oligozoospermic men",
volume = "126",
doi = "10.48101/ujms.v126.7673"
}

Janković, T., Danilović Luković, J., Miler, I., Mitic, N., Hajduković, L.,& Janković, M.. (2021). Assembly of tetraspanins, galectin-3, and distinct N-glycans defines the solubilization signature of seminal prostasomes from normozoospermic and oligozoospermic men. in Upsala journal of medical sciences
Upsala Medical Society., 126.
https://doi.org/10.48101/ujms.v126.7673

Janković T, Danilović Luković J, Miler I, Mitic 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. in Upsala journal of medical sciences. 2021;126.
doi:10.48101/ujms.v126.7673 .

Janković, Tamara, Danilović Luković, Jelena, Miler, Irena, Mitic, Ninoslav, Hajduković, Ljiljana, Janković, Miroslava, "Assembly of tetraspanins, galectin-3, and distinct N-glycans defines the solubilization signature of seminal prostasomes from normozoospermic and oligozoospermic men" in Upsala journal of medical sciences, 126 (2021),
https://doi.org/10.48101/ujms.v126.7673 . .