bims-supasi 2025-02-09 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2025–02–09
twelve papers selected by
Jonathan Wolf Mueller, University of Birmingham

Isolation, structural characterization of natural chondroitin sulfate oligosaccharides and their binding study with anti-angiogenic factors.
Searching for Sulfotyrosines (sY) in a HA(pY)STACK.
A Robust Proteomics-Based Method for Identifying Preferred Protein Targets of Synthetic Glycosaminoglycan Mimetics.
Lactoferrin exhibits PEDV antiviral activity by interfering with spike-heparan sulfate proteoglycans binding and activating mucosal immune response.
Fucoidan alginate and sulfated alginate microbeads induce distinct coagulation, inflammatory and fibrotic responses.
3-Indoleacetic Acid-Modified Chondroitin Sulfate-Mediated Paclitaxel Nanocrystal Assembly for the Treatment of Pancreatic Cancer.
Sulfatase modifying factor 2 as a predictive biomarker for urothelial carcinoma.
Sulfated and Glucuronidated Conjugates of 3-(4-Hydroxy-3-methoxyphenyl) Propionic Acid Can Promote NO Production by Elevated Ca2+ Release from the Endoplasmic Reticulum in HUVECs.
Polyguluronate sulfate has potential to mitigate hyperlipidemia by inhibiting the PCSK9-mediated degradation of LDLR.
Arabidopsis thaliana accumulates dehydroepiandrosterone after infection with phytopathogenic fungi - Effects on plants and fungi.
Understanding the biochemistry of hormones - message in a bottle.
Low-Range Heparin and Protamine Detection: A Single-Center Prospective Diagnostic Study.

Carbohydr Polym. 2025 Apr 01. pii: S0144-8617(25)00043-8. [Epub ahead of print]353 123262

Isolation, structural characterization of natural chondroitin sulfate oligosaccharides and their binding study with anti-angiogenic factors.

Yuxia Liu, Ming Wei, Guo Li, Yilong Zhao, Xiuzhen Yan, Shukai Wang, Xuezheng Song, Zhongfu Wang, Linjuan Huang.

  Drugs that inhibit tumor angiogenesis, promote vascular normalization and improve the tumor microenvironment. However, their application is limited by adaptive or compensatory resistance. Chondroitin sulfate (CS) regulates numerous proteins including pro-angiogenic growth factors, for whom binding affinity depends on sulfation of CS. In this study, we aimed to determine how sulfation of natural tetrasaccharides and hexasaccharides of CS affected binding to the vascular endothelial growth factor (VEGF-A) and fibroblast growth factor 2 (FGF-2). Twenty-eight CS oligosaccharide isomers were obtained by preparative HPLC, tagged with the AEAB fluorescent linker, and identified using an improved chemical derivatization strategy combined with tandem mass spectrometry. CS oligosaccharide microarrays revealed that VEGF-A and FGF-2 bound preferentially to highly sulfated CS, and the GalNAc(4S)GlcA(2S)GalNAc(6S) sequence was found to be indispensable for binding to these proteins. By integrating glycan microarrays with computational modeling, this study revealed the relationship between the structure of CS and its interactions with pro-angiogenic factors. The degree and the specific sulfation patterns on CS should be taken into account when designing anti-angiogenic drugs.

Keywords:  Anti-angiogenesis; Chondroitin sulfate oligosaccharides; Molecular fine-specificities; Natural glycan microarray; Pro-angiogenic growth factors; Structure analysis

DOI:  https://doi.org/10.1016/j.carbpol.2025.123262
J Proteome Res. 2025 Feb 05.

Searching for Sulfotyrosines (sY) in a HA(pY)STACK.

Jordan Tzvetkov, Claire E Eyers, Patrick A Eyers, Kerry A Ramsbottom, Sally O Oswald, John A Harris, Zhi Sun, Eric W Deutsch, Andrew R Jones.

  Protein sulfation can be crucial in regulating protein-protein interactions but remains largely underexplored. Sulfation is nearly isobaric to phosphorylation, making it particularly challenging to investigate using mass spectrometry. The degree to which tyrosine sulfation (sY) is misidentified as phosphorylation (pY) is, thus, an unresolved concern. This study explores the extent of sY misidentification within the human phosphoproteome by distinguishing between sulfation and phosphorylation based on their mass difference. Using Gaussian mixture models (GMMs), we screened ∼45 M peptide-spectrum matches (PSMs) from the PeptideAtlas human phosphoproteome build for peptidoforms with mass error shifts indicative of sulfation. This analysis pinpointed 104 candidate sulfated peptidoforms, backed up by Gene Ontology (GO) terms and custom terms linked to sulfation. False positive filtering by manual annotation resulted in 31 convincing peptidoforms spanning 7 known and 7 novel sY sites. Y47 in calumenin was particularly intriguing since mass error shifts, acidic motif conservation, and MS2 neutral loss patterns characteristic of sulfation provided strong evidence that this site is sulfated rather than phosphorylated. Overall, although misidentification of sulfation in phosphoproteomics data sets derived from cell and tissue intracellular extracts can occur, it appears relatively rare and should not be considered a substantive confounding factor in high-quality phosphoproteomics data sets.

Keywords:  large scale meta-analysis; mass spectrometry; misidentification; phosphorylation; proteomics; tyrosine sulfation

DOI:  https://doi.org/10.1021/acs.jproteome.4c00907
bioRxiv. 2025 Jan 24. pii: 2025.01.23.634492. [Epub ahead of print]

A Robust Proteomics-Based Method for Identifying Preferred Protein Targets of Synthetic Glycosaminoglycan Mimetics.

Daniel K Afosah, Ravikumar Ongolu, Rawan M Fayyad, Adam Hawkridge, Umesh R Desai.

A robust technology is critically needed for identifying preferred protein targets of glycosaminoglycans (GAGs), and synthetic mimetics thereof, in biological milieu. We present a robust 10-step strategy for identification and validation of preferred protein targets of highly sulfated, synthetic, small, GAG-like molecules using diazirine-based photoaffinity labeling- proteomics approach. Our work reveals that optimally designed, homogeneous probes based on minimalistic photoactivation and affinity pulldown groups coupled with rigorous proteomics, biochemical and orthogonal validation steps offer excellent potential to identify preferred targets of GAG mimetics from the potentially numerous possible targets that cloud GAG interaction studies. Application of this 10-step strategy for a promising highly sulfated, small GAG mimetic led to identification of only a handful of preferred targets in human plasma. This new robust strategy will greatly aid drug discovery and development efforts involving GAG sequences, or sulfated small mimetics thereof, as leads.

DOI: https://doi.org/10.1101/2025.01.23.634492
Vet Res. 2025 Jan 31. 56(1): 25

Lactoferrin exhibits PEDV antiviral activity by interfering with spike-heparan sulfate proteoglycans binding and activating mucosal immune response.

Peng Liu, Jinjiao Zuo, Hui Lu, Bin Zhang, Caihong Wu.

  Neonatal piglets infected with Porcine Epidemic Diarrhea Virus (PEDV) experience a mortality rate of up to 90%, resulting in significant economic losses to the swine industry in China. Current strategies using specific antibodies in sow milk to prevent Porcine Epidemic Diarrhea (PED) in these piglets through specific antibodies in sow milk is unsatisfactory. Preliminary studies have shown limited success. Emerging evidence suggests that general immune factors in sow milk provide protection to neonatal piglets, particularly, lactoferrin, play a crucial role in protecting piglets by inhibiting PEDV replication. However, the precise mechanism by which lactoferrin exerts its antiviral effects remains unclear. This study sought to clarify these mechanisms through both in vitro and in vivo approaches, proposing that higher concentrations of lactoferrin lead to greater antiviral activity. It was hypothesized that lactoferrin can impede PEDV by blocking its binding to heparan sulfate proteoglycans (HSPG) on the surface of target cells, and molecular docking experiments was conducted to identify the binding sites between lactoferrin and HSPG. Additionally, the findings indicated that lactoferrin can effectively trigger the maturation of porcine dendritic cells and boosts their antigen-presenting functions, thereby improving intestinal mucosal immunity in neonatal piglets against PEDV. Overall, these findings aid to elucidate the antiviral actions and mechanisms of lactoferrin in sow colostrum, offering new insights for the effective prevention and control of PED in neonatal piglets.

Keywords:  Lactoferrin; PEDV; heparan sulfate proteoglycans; mucosal immunity; neonatal piglets

DOI:  https://doi.org/10.1186/s13567-025-01456-5
Mater Today Bio. 2025 Apr;31 101474

Fucoidan alginate and sulfated alginate microbeads induce distinct coagulation, inflammatory and fibrotic responses.

Kalaiyarasi Vasuthas, Joachim Sebastian Kjesbu, Alessandro Brambilla, Maya Levitan, Abba Elizabeth Coron, Davi M Fonseca, Berit L Strand, Geir Slupphaug, Anne Mari A Rokstad.

  This study investigates the host response to fucoidan alginate microbeads in comparison to sulfated alginate microbeads, which are relevant for immune protection in cell therapy. While sulfated alginate microbeads reduce fibrosis and inflammation, fucoidan, a kelp-derived polysaccharide rich in sulfate groups, has not been evaluated in this context. The study assesses surface reactivity to acute-phase proteins and cytokines using ex vivo human whole blood and plasma models. It also examines pericapsular overgrowth (PFO) in C57BL/6JRj mice, incorporating protein pattern mapping through LC-MS/MS proteomics. Fucoidan alginate microbeads activated complement and coagulation, while both fucoidan and sulfated alginate microbeads induced plasmin activity. Fucoidan alginate microbeads exhibited a distinct cytokine profile, characterized by high levels of MCP-1, IL-8, IFN-γ, and reduced levels of RANTES, Eotaxin, PDGF-BB, TGF-β isoforms, along with higher PFO. The balance between plasmin activity and coagulation emerged as a potential predictor of fibrosis resistance, favouring sulfated alginate microbeads. Explanted materials were enriched with both complement and coagulation activators (Complement C1q and C3, Factor 12, Kallikrein, HMW-kininogen) and inhibitors (C1-inhibitor, Factor H, Factor I). Fucoidan alginate microbeads predominantly enriched extracellular matrix factors (Fibrinogen, Collagen, TGF-β, Bmp), while sulfated alginate microbeads favoured ECM-degrading proteases (Metalloproteases and Cathepsins). This study reveals significant differences in host responses to fucoidan and sulfated alginate in microbeads. The plasmin activity to coagulation ratio is highlighted as a key indicator of fibrosis resistance. Additionally, the preferential enrichment of ECM-degrading proteases on the material surface post-implantation proved to be another crucial factor.

Keywords:  ECM-Degrading proteases; Fibrinolysis; Hydrogels; LC/MS-Proteomics; Sulfated polysaccharides; Surface

DOI:  https://doi.org/10.1016/j.mtbio.2025.101474
ACS Appl Mater Interfaces. 2025 Feb 04.

3-Indoleacetic Acid-Modified Chondroitin Sulfate-Mediated Paclitaxel Nanocrystal Assembly for the Treatment of Pancreatic Cancer.

Yueting Zhu, Aolei Hu, Meilin Chen, Yunting Zhang, Tao Gong, Zhirong Zhang, Ruilian Yu, Yao Fu.

  High drug-loading nanocrystals show significant advantages in delivering hydrophobic small-molecule drugs. However, these nanocrystals face challenges, including inherent instability and limited targetability. In this study, we developed a paclitaxel nanocrystal delivery platform based on chondroitin sulfate modified with 3-indoleacetic acid (CS-IAA). Paclitaxel and CS-IAA assembled into stable nanocrystals (PTX@CS-IAA, PC) with a high drug loading (up to 46.6%), facilitated by π-π stacking and hydrophobic interactions. CS-IAA targets tumors through CD44 receptors, which helps to minimize off-target effects. Additionally, CS-IAA, serving as a functional delivery vehicle, can significantly increase reactive oxygen species (ROS) levels at the tumor site. In an orthotopic pancreatic cancer mouse model, PTX@CS-IAA nanocrystals showed significantly enhanced antitumor effects. When PTX@CS-IAA was combined with the αPD-L1 antibody, the survival of mice with pancreatic tumors was further prolonged. This study provides valuable insights into alternative treatment options for pancreatic cancer, with the potential to improve patient prognosis and survival.

Keywords:  3-indoleacetic acid; chondroitin sulfate; nanocrystal; paclitaxel; pancreatic cancer; αPD-L1 antibody; π−π stacking

DOI:  https://doi.org/10.1021/acsami.4c19450
Discov Oncol. 2025 Feb 07. 16(1): 126

Sulfatase modifying factor 2 as a predictive biomarker for urothelial carcinoma.

Wei-Ting Kuo, Yi-Chen Lee, Jia-Bin Liao, Ching-Jiunn Tseng, Yi-Fang Yang.

  Sulfatases mediate the sulfation level of cell-surface heparan sulfates to regulate signal transduction, thereby promoting cancer progression. Sulfatase activation requires a sulfatase modifying factor (SUMF) for the modification of its catalytic domain. The role of the SUMF family in urothelial carcinoma (UC) has not been adequately evaluated. In this study, we used an online database and immunohistochemistry to assess genetic changes and the mRNA and protein expression of SUMFs and related candidate targets in UC. We found that SUMF1 and SUMF2 were amplified in UC tissues. High SUMF2 mRNA levels were associated with poor overall survival (OS) and disease-free survival (DFS) in bladder UC (BLCA) from The Cancer Genome Atlas (TCGA) dataset. High SUMF2 protein levels were associated with grade (P < 0.001), T status (P = 0.01), and stage (P = 0.006) in patients with BLCA. We also examined SUMF2 expression levels in upper tract UC (UTUC). SUMF2 expression was associated with stage (P = 0.046), poor OS (P = 0.0022), and DFS (P = 0.019) in patients with UTUC. Knockdown of SUMF2 significantly reduced the migration and invasion abilities of 5637 cells. Furthermore, SUMF2 mRNA levels negatively correlated with FBXW7 mRNA levels in BLCA. The SUMF2high/FBXW7low expression profile predicted the worst survival in BLCA. Taken together, SUMF2 expression is linked to unfavorable clinical outcomes in patients with UC and may serve as a useful prognostic biomarker for UC staging.

Keywords:  FBXW7; SUMF2; Sulfatases; Urothelial carcinoma

DOI:  https://doi.org/10.1007/s12672-025-01859-y
ACS Omega. 2025 Jan 28. 10(3): 2887-2896

Sulfated and Glucuronidated Conjugates of 3-(4-Hydroxy-3-methoxyphenyl) Propionic Acid Can Promote NO Production by Elevated Ca2+ Release from the Endoplasmic Reticulum in HUVECs.

Tint Ni Ni Tun, Susumu Yoshino, Hiroyuki Kayaki, Hiroshige Kuwahara, Toshiro Matsui.

We aimed to clarify whether metabolic conjugates of sulfated and glucuronidated forms have the physiological potential to produce the vasorelaxant nitric oxide (NO) in human umbilical vein endothelial cells (HUVECs), using 3-(4-hydroxy-3-methoxyphenyl) propionic acid (HMPA), a metabolite of dietary flavonoids in the gut. Treatment of HUVECs with sulfated and glucuronidated HMPAs significantly increased NO production and eNOS phosphorylation. A transporter-inhibitor-aided cellular uptake experiment of HMPAs revealed that both conjugates were incorporated into cells via MCT, OATP1A2, and GLUT transporters, whereas intact HMPA was transported via the MCT and OATP1A2 routes. A Fluo-4-probe Ca2+ assay demonstrated that the incorporated HMPAs significantly increased intracellular Ca2+ concentration by stimulating the IP3R of the endoplasmic reticulum in the CaMKII/eNOS signaling cascade. In conclusion, to our knowledge, this study provides the first evidence that sulfated and glucuronidated forms of HMPAs may stimulate NO production in HUVECs.

DOI: https://doi.org/10.1021/acsomega.4c09008
Int J Biol Macromol. 2025 Feb 02. pii: S0141-8130(25)01134-1. [Epub ahead of print]302 140585

Polyguluronate sulfate has potential to mitigate hyperlipidemia by inhibiting the PCSK9-mediated degradation of LDLR.

Dan Li, Meijie Xu, Dingfu Wang, Shixin Wang, Chunxia Li.

  Hyperlipidemia has become a major global health challenge and one of the leading causes of mortality. Proprotein convertase subtilisin/kexin type 9 (PCSK9), a circulating plasma protein, promotes the lysosomal degradation of hepatic low-density lipoprotein receptors (LDLR), thereby reducing the clearance of low-density lipoprotein (LDL) from blood plasma. PCSK9 has increasingly become a prominent therapeutic target for the development of lipid-lowering agents. In this study, we firstly identified polyguluronate sulfate (PGS) as a novel PCSK9 inhibitor. PGS could bind to the positively charged domain of PCSK9 with a KD value of 3.198 μM, effectively blocking its mediated LDLR degradation. This interaction leads to increase in LDLR levels on hepatocyte surface, enhancing LDL clearance. Furthermore, we demonstrated that PGS more effectively activates the AMP-activated protein kinase (AMPK) pathway compared to polymannuronate sulfate (PMS) at 200 μg/mL, resulting in about 2-fold greater lipid-lowering effect. In summary, our findings highlight PGS as a promising candidate for the development of novel lipid-lowering drugs, offering new insights into the therapeutic potential of sulfate polysaccharides targeting PCSK9.

Keywords:  Hyperlipidemia; PCSK9 inhibitor; Polyguluronate sulfate

DOI:  https://doi.org/10.1016/j.ijbiomac.2025.140585
Plant Physiol Biochem. 2025 Jan 28. pii: S0981-9428(25)00098-1. [Epub ahead of print]221 109570

Arabidopsis thaliana accumulates dehydroepiandrosterone after infection with phytopathogenic fungi - Effects on plants and fungi.

Ceren Oktay, Glendis Shiko, Maximilian Liebl, Felix Feistel, Sarah Mußbach, Karl Ludwig Körber, Emanuel Barth, Ludwig Huber, Anna Antony, Ralf Oelmüller, Michael Reichelt, Kilian Ossetek, Christoph Müller, Alexandra C U Furch, Jan Klein.

  Progestogens and androgens have been found in many plants, but little is known about their physiological function. We used a previously established UPLC-ESI-MS/MS method to analyze progestogen and androgen profiles in fungal infections. Here we show that dehydroepiandrosterone (DHEA), a C19 steroid, specifically accumulates in shoots of Arabidopsis thaliana (L.) HEYNH. infected with Alternaria brassicicola (SCHWEIN.) WILTSHIRE. Elevated DHEA levels in plants seem not to be product of fungal sterol/steroid precursor activity, but an intrinsic plant response to the infection. DHEA was applied exogenously to analyze the effects of the androgen on development and gene expression in A. thaliana. Our findings reveal that DHEA treatment downregulates membrane-associated, salicylic acid and abscisic acid-regulated, as well as stress-responsive genes. Notably, DHEA does not inhibit the isoprenoid or post-lanosterol pathway of the ergosterol biosynthesis. Moreover, A. brassicicola was also treated with DHEA to analyze the growth, sterol pattern and membrane-integrity. Our data suggest that DHEA enhances the permeability of plant and fungal biomembranes. We propose that DHEA accumulation is a plant defense response which reduces fungal growth in plant tissues.

Keywords:  Alternaria brassicicola; Arabidopsis thaliana; Cell membrane integrity; Dehydroepiandrosterone; Ergosterol biosynthesis; Fungal infection; Mammalian steroids

DOI:  https://doi.org/10.1016/j.plaphy.2025.109570
Essays Biochem. 2025 Feb 06. 69(1): 1-18

Understanding the biochemistry of hormones - message in a bottle.

Dominic C Y Lai, Jonathan Wolf Mueller.

  Hormones play pivotal roles in our well-being, and even more so in times of stress or disease. They determine body composition and govern reproductive processes. Hormonal compounds tend to be evolutionarily very old compounds, but only coevolved receptor systems make up powerful biological signals. We will discuss what makes some metabolites good building materials for hormones and how information may be encoded, using these scaffolds. Starting with hormone biosynthesis and regulated release from secreting cells, we will look at different stages of the whole hormone signaling process: the distribution of the hormonal "message-in-a-bottle" throughout the body, the passing of some hormones through membranes, and pre-receptor metabolism. Binding to different classes of receptors is not the end of hormone signaling, but the beginning of a second phase of signaling via second messengers, before hormonal messages are switched off again. Studying hormone biochemistry will produce exciting new findings in the future.

Keywords:  anthropomorphic storytelling; anthropomorphism; difficult-to-teach subject; innovative teaching material

DOI:  https://doi.org/10.1042/EBC20240039
J Cardiothorac Vasc Anesth. 2025 Jan 17. pii: S1053-0770(25)00038-2. [Epub ahead of print]

Low-Range Heparin and Protamine Detection: A Single-Center Prospective Diagnostic Study.

Michael Vandenheuvel, Laura Vierstraete, Filip De Somer, Katrien M J Devreese, Patrick F Wouters, Pieter M De Kesel.

   OBJECTIVES: The detection of low-range heparin activity is important to correctly assess heparin reversal and rebound, especially after cardiopulmonary bypass. Current parameters are either not available at point-of-care (anti-Xa activity [aXa] and activated partial thromboplastin time [aPTT]), insensitive (kaolin-activated clotting time [kACT]), or expensive (ROTEM viscoelastic test). We aimed to assess the performance of a recently proposed parameter from the Sonoclot viscoelastic test: the slope-45. We aimed to assess the effects of a range of low-dose heparin and protamine and their interaction on multiple proposed parameters.
DESIGN: Prospective in vitro volunteer study.
SETTING: Single-center university teaching hospital.
PARTICIPANTS: Healthy volunteers.
INTERVENTIONS: Blood samples from healthy volunteers (n = 10) were treated ex vivo with incremental low doses of heparin, protamine, or their combination. In the combination cycle, theoretical reversal ratios were 25%, 62.5%, 100%, and 200% of the highest heparin dose, based on a unit-for-unit reversal. We compared the effects on aXa, aPTT, iSTAT kACT, ROTEM clotting time ratio, and the novel Sonoclot slope-45 parameter, and we performed receiver operating curve analysis.
MEASUREMENTS AND MAIN RESULTS: In heparin-spiked blood, all parameters except iSTAT kACT were able to reliably detect low heparin activity. Protamine, both in isolation and combined with heparin, showed no impact on aXa, aPTT, iSTAT kACT, ROTEM clotting time ratio, and the novel Sonoclot slope-45 parameter.
CONCLUSIONS: We were able to confirm that the Sonoclot slope-45 has a high sensitivity for low-dose heparin, which is retained in the setting of interacting protamine. It was insensitive to protamine in itself.

Keywords:  blood coagulation tests; heparin; point-of-care testing; protamine; viscoelastic testing

DOI:  https://doi.org/10.1053/j.jvca.2025.01.009