A novel pyruvate kinase (PK-S) from boar spermatozoa is localized at the fibrous sheath and the acrosome

Feiden, Sandra; Stypa, Heike; Wolfrum, Uwe; Wegener, G.; Kamp, Günter

doi:10.1530/rep-06-0250

Cited by 23 publications

(34 citation statements)

References 53 publications

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“…A defective PKM 2 has the potential to create an imbalance in the equilibrium in growth and apoptosis (45,46) thus resulting in growth retardation in BS. Similarly, a defective sperm-specific pyruvate kinase would be unable to maintain normal glycolysis, crucial for sperm mobility (47,48), to explain male-specific infertility in BS. Also, PKM 2 has been reported to interact with microtubules physically (49), with the possibility to affect the motility of microtubule-rich cells like sperm cells.…”

Section: Discussionmentioning

confidence: 99%

Dominant Negative Mutations Affect Oligomerization of Human Pyruvate Kinase M2 Isozyme and Promote Cellular Growth and Polyploidy

Gupta

Kalaiarasan

Faheem

et al. 2010

Journal of Biological Chemistry

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This study was designed to understand the mechanism and functional implication of the two heterozygous mutations (H391Y and K422R) of human pyruvate kinase M2 isozyme (PKM 2 ) observed earlier in a Bloom syndrome background. The co-expression of homotetrameric wild type and mutant PKM 2 in the cellular milieu resulting in the interaction between the two at the monomer level was substantiated further by in vitro experiments. The cross-monomer interaction significantly altered the oligomeric state of PKM 2 by favoring dimerization and heterotetramerization. In silico study provided an added support in showing that hetero-oligomerization was energetically favorable. The hetero-oligomeric populations of PKM 2 showed altered activity and affinity, and their expression resulted in an increased growth rate of Escherichia coli as well as mammalian cells, along with an increased rate of polyploidy. These features are known to be essential to tumor progression. This study provides insight in understanding the modulated role of large oligomeric multifunctional proteins such as PKM 2 by affecting cellular behavior, which is an essential observation to understand tumor sustenance and progression and to design therapeutic intervention in future.A variety of genetic diseases and experimental situations within the heterozygous state depict an allelic relationship where the mutant recessive allele overrides the function of its normal (wild type) dominant allele. This condition, referred to as dominant negative, is usually observed in the case of oligomeric or multidomain proteins with a possibility of cross-monomer interaction. Such mutant proteins by acting as competitive inhibitors of the normal protein function could generate polymorphic forms in a single cell. A phenomenon observed in collagen, where dominant negative mutations cause the production of abnormal oligomers (1, 2), and in transcription factors like helix-loop-helix and leucine zippers, where mutant monomers sequestering the function of wild type in a dimer bound to DNA, leads to an altered gene expression (3-5). Dominant negative mutations are also reported to affect some multifunctional molecules like p53 with differential impact on cell physiology (6 -10).Pyruvate kinase (EC 2.7.1.40) catalyzes irreversibly the transphosphorylation from P-enolpyruvate to ADP-generating pyruvate and ATP in glycolysis (11,12). Depending upon the differential metabolic requirements of the tissues, the enzyme is expressed in four different isoforms, L, R, M 1 , and M 2 in vertebrates (13). PKM 2 2 is a ubiquitous, prototype enzyme, present in all tissues during embryonic stage, and is gradually replaced by other isozymic forms in specific tissues, during development. It is necessary for cellular division irrespective of the type of tissue and reappears during cellular division and tumor formation (14 -17). PKM 2 is known to regulate its activity by switching between an active tetramer and inactive dimer form in a fructose 1,6-bisphosphate-dependent manner to shift the cellular met...

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Section: Discussionmentioning

confidence: 99%

Dominant Negative Mutations Affect Oligomerization of Human Pyruvate Kinase M2 Isozyme and Promote Cellular Growth and Polyploidy

Gupta

Kalaiarasan

Faheem

et al. 2010

Journal of Biological Chemistry

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“…"Aerobic glycolysis" is known to occur in erythrocytes because these cells do not contain mitochondria [36]. It is also described for sperm where the mitochondria are compartmentalized in the short midpiece of the flagellum and glycolysis may supply the mitochondrial-free region with ATP [37]. Warburg et al observed "aerobic glycolysis" in various tumours (Warburg effect) and believed that an impairment of mitochondrial function caused "aerobic glycolysis" [38].…”

Section: Discussionmentioning

confidence: 99%

Changes in Human Endothelial Cell Energy Metabolic Capacities during in vitro Cultivation. The Role of “Aerobic Glycolysis” and Proliferation

Peters

Kamp²,

Berz³

et al. 2009

Cell Physiol Biochem

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Background: In this study the influence of cultivation and proliferation on energy metabolic characteristics of human umbilical vein endothelial cells (HUVEC) has been examined. The energy metabolic capacities of human endothelial cells freshly isolated from the umbilical vein were compared with those after cultivation for three passages and as subconfluent and confluent cultures. Methods: Expression of cell type-specific differentiation markers and proliferative activity were studied in dependency on cultivation characteristics. Furthermore, the energy metabolic characteristics of HUVEC were analyzed by measurement of the maximum catalytic activities of marker enzymes of various metabolic pathways. Results: Examination of a typical marker of proliferation, Ki67, confirmed that HUVEC changed in culture from a non-proliferative to a proliferative state. Compared to other cell types, the enzyme pattern of HUVEC showed a high glycolytic and a high NADPH regenerating capacity. These capacities increased by cultivation nearly to the same degree as marker enzymes of other metabolic pathways (e.g. citric acid cycle). Conclusion: Our data support the theory that metabolism of EC is primarily by “aerobic glycolysis”, i.e. the conversion of glucose to lactate in the presence of oxygen. These characteristics were independent of whether the cells are freshly isolated/non-proliferating or cell culture-adapted/proliferating.

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“…Interestingly, we found that PK was localized in the acrosomal area and the midpiece, while LDH was expressed in the tail. The observations were in according with Feiden et al [36] in boar sperm and Burgos et al [37] in mouse sperm. Exposure goat sperm to AICAR or Met enhanced activity of PK and LDH, while Compound C inhibited activity of both PK and LDH ( Fig.…”

Section: Cellular Physiology and Biochemistrymentioning

confidence: 95%

5’-AMP-Activated Protein Kinase Regulates Goat Sperm Functions via Energy Metabolism In Vitro

Zhu

et al. 2018

Cell Physiol Biochem

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Background/Aims: ATP is essential for mammalian sperm to survive and maintain fertilizing capacity. AMP-activated protein kinase (AMPK) is a sensor of cellular energy status. The aims of the present study were to explore the localization of AMPK in goat sperm and to investigate whether and how AMPK regulates sperm functions in vitro. Methods: Sperm were treated with AMPK modulators (AICAR, metformin and Compound C) during incubation. Sperm motility was assessed with a computer-assisted spermatozoa analysis system (CASA). Membrane integrity, acrosome reaction and mitochondrial membrane potentials were detected by SYBR-14/PI, FITC-PNA and JC-1 staining, respectively. And the lactate content, ATP content, AMPK activity, activity of pyruvate kinase (PK) and lactate dehydrogenase (LDH) were also measured with the commercial assay kits. Immunofluorescence staining was used to analyze the distribution of PK, LDH, AMPK and phospho-Thr172-AMPK in sperm. The role of AMPK was further studied during induction of capacitation and acrosome reaction. Results: We found that AMPKα was localized in the entire acrosomal region, the midpiece and the flagellum, while the phospho-Thr172-AMPK was distributed in the head, the midpiece and flagellum. Activation of AMPK by AICAR and metformin significantly improved sperm motility, membrane integrity and acrosome reaction, largely maintained sperm mitochondrial membrane potentials, lactate content and ATP content, and enhanced the activity of AMPK, PK and LDH, whereas inhibition by Compound C triggered the converse effects. Moreover, PK was localized in the acrosomal area and the midpiece, while LDH was distributed in the tail. Induction of capacitation and acrosome reaction led to AMPK phosphorylation. AMPK phosphorylation regulated the activity of energetic enzymes. Conclusion: This study for the first time provides evidence that AMPK governs goat sperm functions through energy metabolism in vitro. This finding will help to improve assisted reproductive techniques in goats and the other species.

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A novel pyruvate kinase (PK-S) from boar spermatozoa is localized at the fibrous sheath and the acrosome

Cited by 23 publications

References 53 publications

Dominant Negative Mutations Affect Oligomerization of Human Pyruvate Kinase M2 Isozyme and Promote Cellular Growth and Polyploidy

Dominant Negative Mutations Affect Oligomerization of Human Pyruvate Kinase M2 Isozyme and Promote Cellular Growth and Polyploidy

Changes in Human Endothelial Cell Energy Metabolic Capacities during in vitro Cultivation. The Role of “Aerobic Glycolysis” and Proliferation

5’-AMP-Activated Protein Kinase Regulates Goat Sperm Functions via Energy Metabolism In Vitro

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