Conserved core tryptophans of FnII domains are crucial for the membranolytic and chaperone‐like activities of bovine seminal plasma protein PDC‐109

Singh, Bhanu; Asthana, Abhishek; Basu, Amrita; Tangirala, Ramakrishna; Rao, Chintalagiri Mohan; Swamy, Musti J.

doi:10.1002/1873-3468.13617

Cited by 5 publications

(3 citation statements)

References 37 publications

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“…Additionally, we also identified the differential seminal plasma protein PDC-109, which contains two fibronectin type II (FnII) structural domains that aid in sperm capacitation by binding to choline phospholipids on the sperm plasma membrane to induce lipid efflux; this, in turn, allows for successful sperm fertilization. In addition, PDC-109, like HSP90AA1, displays chaperone-like properties that protect other proteins from various types of stress [51], protecting the functional status of sperm from harmful environmental factors, such as freezing and AI, on male reproductive function.…”

Section: Differences In Spermatozoa Proteins Between Bucks and Ramsmentioning

confidence: 99%

TMT-Based Comparative Proteomic Analysis of the Spermatozoa of Buck (Capra hircus) and Ram (Ovis aries)

Ren

Chen

Tang

et al. 2023

Genes

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Spermatozoa are unique cells that carry a library of proteins that regulate the functions of molecules to achieve functional capabilities. Currently, large amounts of protein have been identified in spermatozoa from different species using proteomic approaches. However, the proteome characteristics and regulatory mechanisms of spermatozoa in bucks versus rams have not been fully unraveled. In this study, we performed a tandem mass tag (TMT)-labeled quantitative proteomic analysis to investigate the protein profiles in the spermatozoa of buck (Capra hircus) and ram (Ovis aries), two important economic livestock species with different fertility potentials. Overall, 2644 proteins were identified and quantified via this approach. Thus, 279 differentially abundant proteins (DAPs) were filtered with a p-value < 0.05, and a quantitative ratio of >2.0 or <0.5 (fold change, FC) in bucks versus rams, wherein 153 were upregulated and 126 were downregulated. Bioinformatics analysis revealed that these DAPs were mainly localized in the mitochondria, extracellular and in the nucleus, and were involved in sperm motility, membrane components, oxidoreductase activity, endopeptidase complex and proteasome-mediated ubiquitin-dependent protein catabolism. Specifically, partial DAPs, such as heat shock protein 90 α family class a member 1 (HSP90AA1), adenosine triphosphate citrate lyase (ACLY), proteasome 26S subunit and non-ATPase 4 (PSMD4), act as “cross-talk” nodes in protein–protein networks as key intermediates or enzymes, which are mainly involved in responses to stimuli, catalytic activity and molecular function regulator pathways that are strictly related to spermatozoa function. The results of our study offer valuable insights into the molecular mechanisms of ram spermatozoa function, and also promote an efficient spermatozoa utilization link to fertility or specific biotechnologies for bucks and rams.

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Section: Differences In Spermatozoa Proteins Between Bucks and Ramsmentioning

confidence: 99%

TMT-Based Comparative Proteomic Analysis of the Spermatozoa of Buck (Capra hircus) and Ram (Ovis aries)

Ren

Chen

Tang

et al. 2023

Genes

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“…Proteínas como la Espermadhesina-1 (I. Bustamante-Filho et al, 2014) y la Binder Sperm Protein (BSP) (Singh et al, 2020) han sido producidas de forma heteróloga usando E. coli como fábrica celular, con resultados interesantes en cuanto a la estructura y función de estas proteínas y sobre su uso para mejorar la calidad del semen en procesos de biotecnología reproductiva. De acuerdo con los perfiles de expresión, tanto la Espermadhesina como la BSP tienen tendencia a formar agregados insolubles en forma de cuerpos de inclusión por la presencia de múltiples enlaces disulfuro, lo que puede limitar su aplicación práctica.…”

Section: Introductionunclassified

Producción Y Purificación De Osteopontina Bovina Recombinante Mediante Escherichia coli Como Fábrica Celular

et al. 2022

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Los programas de reproducción y mejoramiento animal requieren la optimización de herramientas biotecnológicas capaces de favorecer los índices reproductivos en diversas especies. El uso de aditivos proteicos que mejoren la criopreservación espermática y la producción de embriones in vitro, parece ser una alternativa interesante. La Osteopontina se ha relacionado con el potencial fecundante del espermatozoide y con el desarrollo embrionario temprano. El objetivo de este trabajo fue determinar las condiciones óptimas para la producción de Osteopontina recombinante (rOPN) mediante el uso de Escherichia coli como fábrica celular. Para esto, el gen de la OPN se insertó en un vector de expresión pET28(a+) inducible por IPTG, con resistencia a la Kanamicina y una cola de histidinas (6xHis-tag). El constructo resultante se usó para transformar células competentes de E. Coli BL21-Star TM. Las colonias transformadas se usaron para la producción de rOPN-H6 a 20, 30 y 37 °C, probándose dos concentraciones del inductor IPTG (1.0 y 0.1mM). Se realizó una purificación de la rOPN-H6 mediante columnas de afinidad con imidazol (10, 50, 200, 350, 500mM). Los resultados evidenciaron que la producción de rOPN-H6 solo fue exitosa a 37°C independiente de la concentración de IPTG empleada. La purificación de la rOPN-H6 fue exitosa usando imidazol a 200mM, con una aparente tendencia a la dimerización luego de obtener la proteína purificada. De este modo, se concluye cuáles son las mejores condiciones para obtener la OPN recombinante, sugiriendo su potencial uso en ensayos de criopreservación espermática y en medios de cultivo para producción de embriones in vitro.

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“…Indeed, the heterocyclic ring of tryptophan exhibits a wide spectrum of polarity that has been demonstrated to interact with various components of the lipid membrane domains 2 . It can associate with a cationic group such as choline through π-based interactions 3,4 , the hydrophobic alkyl chain of a fatty acid or amino acid [5][6][7] , or a H-bond acceptor such as the phosphate head group by donating the indole's N-H 8,9 . In line with the dynamical nature of proteins 10 , a tryptophan residue may have multiple interacting partners that interchange during the course of action 2 .…”

mentioning

confidence: 99%

Roles of inter- and intramolecular tryptophan interactions in membrane-active proteins revealed by racemic protein crystallography

Lander

Mercado

et al. 2023

Commun Chem

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Tryptophan is frequently found on the surface of membrane-associated proteins that interact with the lipid membrane. However, because of their multifaceted interactions, it is difficult to pinpoint the structure-activity relationship of each tryptophan residue. Here, we describe the use of racemic protein crystallography to probe dedicated tryptophan interactions of a model tryptophan-rich bacteriocin aureocin A53 (AucA) by inclusion and/or exclusion of potential ligands. In the presence of tetrahedral anions that are isosteric to the head group of phospholipids, distinct tryptophan H-bond networks were revealed. H-bond donation by W40 was critical for antibacterial activity, as its substitution by 1-methyltryptophan resulted in substantial loss of activity against bacterial clinical isolates. Meanwhile, exclusion of tetrahedral ions revealed that W3 partakes in formation of a dimeric interface, thus suggesting that AucA is dimeric in solution and dissociated to interact with the phosphate head group in the presence of the lipid membrane. Based on these findings, we could predict the tryptophan residue responsible for activity as well as the oligomeric state of a distant homologue lacticin Q (48%).

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Conserved core tryptophans of FnII domains are crucial for the membranolytic and chaperone‐like activities of bovine seminal plasma protein PDC‐109

Cited by 5 publications

References 37 publications

TMT-Based Comparative Proteomic Analysis of the Spermatozoa of Buck (Capra hircus) and Ram (Ovis aries)

TMT-Based Comparative Proteomic Analysis of the Spermatozoa of Buck (Capra hircus) and Ram (Ovis aries)

Producción Y Purificación De Osteopontina Bovina Recombinante Mediante Escherichia coli Como Fábrica Celular

Roles of inter- and intramolecular tryptophan interactions in membrane-active proteins revealed by racemic protein crystallography

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