bims-supasi 2023-11-19 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2023–11–19
twelve papers selected by
Jonathan Wolf Mueller, University of Birmingham

Identification and structural characterization of a novel chondroitin sulfate-specific carbohydrate-binding module: The first member of a new family, CBMxx.
Restoring the barrier of chronically damaged urothelium using chondroitin sulfate glycosaminoglycan-replenishment therapy: a preclinical study using a chronic experimental model for bladder pain syndrome/interstitial cystitis and reflections on lately published similar models.
The Relation of Serum Dehydroepiandrosterone Sulfate Level to Protein Energy Wasting in Hemodialysis Patients.
Composition and Anticoagulant Potential of Chondroitin Sulfate and Dermatan Sulfate from Inedible Parts of Garfish (Belone belone).
Heparan-6-O-Endosulfatase 2 Promotes Invasiveness of Head and Neck Squamous Carcinoma Cell Lines in Co-Cultures with Cancer-Associated Fibroblasts.
Estrogen Sulfotransferase and Sulfatase in Steroid Homeostasis, Metabolic Disease, and Cancer.
On-demand chlorine dioxide solution enhances odontoblast differentiation through desulfation of cell surface heparan sulfate proteoglycan and subsequent activation of canonical Wnt signaling.
The role of genetic polymorphisms in the sulfation of pregnenolone by human cytosolic sulfotransferase SULT2B1a.
Sex steroid levels in corresponding cerebrospinal fluid and serum samples quantified by mass spectrometry in men.
Galectin-14 promotes hepatocellular carcinoma tumor growth via enhancing heparan sulfate proteoglycan modification.
Insights into the molecular mechanism of dextran sulfate inhibiting DNA digestion by pepsin: a crucial role of sulfate group on the binding to DNA.
Octanoyl esterification of low molecular weight sulfated galactan enhances the cellular uptake and collagen expression in fibroblast cells.

Int J Biol Macromol. 2023 Nov 09. pii: S0141-8130(23)04858-4. [Epub ahead of print] 127959

Identification and structural characterization of a novel chondroitin sulfate-specific carbohydrate-binding module: The first member of a new family, CBMxx.

Guanchen Liu, Yaoguang Chang, Xuanwei Mei, Guangning Chen, Yuying Zhang, Xiaoxiao Jiang, Wenwen Tao, Changhu Xue.

  Chondroitin sulfate is a biologically and commercially important polysaccharide with a variety of applications. Carbohydrate-binding module (CBM) is an important class of carbohydrate-binding protein, which could be utilized as a promising tool for the applications of polysaccharides. In the present study, an unknown function domain was explored from a putative chondroitin sulfate lyase in PL29 family. Recombinant PhCBMxx demonstrated binding capacity to chondroitin sulfates with Ka values of 2.1 ± 0.2 × 106 M-1 and 6.0 ± 0.1 × 106 M-1 to chondroitin sulfate A and chondroitin sulfate C, respectively. The 1.55 Å resolution X-ray crystal structure of PhCBMxx exhibited a β-sandwich structure formed by two antiparallel β-sheets. A binding groove in PhCBMxx interacting with chondroitin sulfate was subsequently identified, and the potential of PhCBMxx for visualization of chondroitin sulfate was evaluated. PhCBMxx is the first characterized chondroitin sulfate-specific CBM. The novelty of PhCBMxx proposed a new CBM family of CBMxx.

Keywords:  Carbohydrate-binding module; Chondroitin sulfate; Crystal structure

DOI:  https://doi.org/10.1016/j.ijbiomac.2023.127959
Curr Opin Urol. 2023 Nov 14.

Restoring the barrier of chronically damaged urothelium using chondroitin sulfate glycosaminoglycan-replenishment therapy: a preclinical study using a chronic experimental model for bladder pain syndrome/interstitial cystitis and reflections on lately published similar models.

Boy B Rozenberg, Charlotte J van Ginkel, Dick A W Janssen.

PURPOSE OF REVIEW: This study aims to further understand the physiological mechanism of chondroitin sulfate treatment on the urinary bladder in cases of inflammation, by investigating the effect of chondroitin sulfate therapy on recovery of urothelial barrier in an in-vitro chronic injury model.
RECENT FINDINGS: With inflammatory bladder conditions, the urothelial barrier seems decreased. Glycosaminoglycan (GAG) replacement therapy is supposed to help restore this barrier. Clinical studies on inflammatory bladder conditions are complicated because of the heterogeneous patient population, hence the need for preclinical models.
SUMMARY: In a model using porcine urothelial cells, functional barrier (TEER) and barrier markers were assessed. Chronic urothelial damage was simulated through protamine sulfate instillations with and without subsequent chondroitin sulfate instillations during 3 days. Chondroitin sulfate instillations significantly improved TEER compared to protamine sulfate treatment only (TEER difference 310 Ω.cm2, P < 0.001). This consistent effect over 3 days resulted in a significant higher mean TEER value in the chondroitin sulfate treated group (difference 1855 Ω.cm2, P < 0.001). Enhanced recovery of chondroitin sulfate and other barrier markers was observed.Chondroitin sulfate therapy shows promise in facilitating the recovery of the urothelial barrier in cases of chronic damage. This preclinical study lends support to the use of clinical GAG replenishment therapy for patients with a chronically impaired urothelium.

DOI: https://doi.org/10.1097/MOU.0000000000001149
Saudi J Kidney Dis Transpl. 2022 Sep 01. 33(5): 639-649

The Relation of Serum Dehydroepiandrosterone Sulfate Level to Protein Energy Wasting in Hemodialysis Patients.

Mohammed Kamal Nassar, Ghada El Kannishy, Eman Nagy, Rasha Samir Shemies, Mostafa Mansour, Mohamed Mofreh, Tamer Zaki Gaber, Samar Tharwat.

Protein-energy wasting (PEW) is a major risk for morbidity and mortality in hemodialysis (HD) patients. The change in the concentration of dehydroepiandrosterone sulfate (DHEA-S) may play a role in PEW. The aim of this work was to study the possible relationship between serum DHEA-S levels and various nutritional and inflammatory parameters in a cohort of HD patients. In total, 78 HD patients (47 males and 31 females) were included in this crosssectional observational study. In addition to taking their history, clinical examinations, and routine laboratory investigations, the nutritional status was assessed, and their serum DHEA-S was measured. Nutritional status was assessed by anthropometric measures, bioelectrical impedance analysis, malnutrition inflammation scores, and subjective global assessments. A diagnosis of malnutrition was made based on the recommendations of the International Society of Renal Nutrition and Metabolism. The relationship between DHEA-S and various nutritional parameters was analyzed. Eighteen patients (23.1%) suffered from PEW. Those with PEW had a longer duration of HD (P = 0.04), and lower serum levels of creatinine (P = 0.003), hemoglobin (P = 0.01), albumin (P <0.0001), cholesterol (P = 0.02), and DHEA-S (P = 0.01). Among the variables, serum DHEA-S levels were significant predictors of PEW in this cohort (odds ratio: 0.976; 95% confidence interval: 0.954-1.0; P = 0.04). PEW is frequently encountered in HD patients. Decreased serum DHEA-S levels were associated with PEW in male HD patients. Further studies are needed to assess the effect of hormone supplementation on this serious disorder in HD patients.

DOI: https://doi.org/10.4103/1319-2442.389424
Foods. 2023 Oct 24. pii: 3887. [Epub ahead of print]12(21):

Composition and Anticoagulant Potential of Chondroitin Sulfate and Dermatan Sulfate from Inedible Parts of Garfish (Belone belone).

Sawssen Ben Chikha, Hajer Bougatef, Federica Capitani, Ikram Ben Amor, Francesca Maccari, Jalel Gargouri, Assaad Sila, Nicola Volpi, Ali Bougatef.

  Glycosaminoglycans (GAGs) play a crucial role due to their significant biomedical functions. Chondroitin sulfate (CS) and dermatan sulfate (DS), the main representative family of GAGs, were extracted and purified from garfish (Belone belone) by-products, i.e., skin (GSB), bones (GCB), and heads (GHB), and their composition and anticoagulant activity were investigated. CS/DS were purified by ion-exchange chromatography with yields of 8.1% for heads, 3.7% for skin, and 1.4% for bones. Cellulose acetate electrophoresis was also explored for analyzing the extracted CS/DS. Interestingly, GHB, GSB, and GCB possessed sulfate contents of 21 ± 2%, 20 ± 1%, and 20 ± 1.5%, respectively. Physico-chemical analysis showed that there were no significant differences (p > 0.05) between the variances for sulfate, uronic acid, and total sugars in the GAGs extracted from the different parts of fish. Disaccharide analysis by SAX-HPLC showed that the GSB and GCB were predominately composed of ΔDi-4S [ΔUA-GalNAc 6S] (74.78% and 69.22%, respectively) and ΔDi-2,4S [ΔUA2S-GalNAc 4S] (10.92% and 6.55%, respectively). However, the GHB consisted of 25.55% ΔDi-6S [ΔUA-GalNAc 6S] and 6.28% ΔDi-2,6S [ΔUA2S-GalNAc 4S]. Moreover, classical anticoagulation tests were also used to measure their anticoagulant properties in vitro, which included the activated partial thromboplastin time, prothrombin time, and thrombin time. The CS/DS isolated from garfish by-products exhibited potent anticoagulant effects. The purified CS/DS showed exceptional anticoagulant properties according to this research and can be considered as a new agent with anticoagulant properties.

Keywords:  Belone belone; anticoagulant activity; by-products; chemical characterization; chondroitine sulfate; dermatan sulfate

DOI:  https://doi.org/10.3390/foods12213887
Cancers (Basel). 2023 Oct 27. pii: 5168. [Epub ahead of print]15(21):

Heparan-6-O-Endosulfatase 2 Promotes Invasiveness of Head and Neck Squamous Carcinoma Cell Lines in Co-Cultures with Cancer-Associated Fibroblasts.

Pritha Mukherjee, Xin Zhou, Julius Benicky, Aswini Panigrahi, Reem Aljuhani, Jian Liu, Laurie Ailles, Vitor H Pomin, Zhangjie Wang, Radoslav Goldman.

  Local invasiveness of head and neck squamous cell carcinoma (HNSCC) is a complex phenomenon supported by interaction of the cancer cells with the tumor microenvironment (TME). We and others have shown that cancer-associated fibroblasts (CAFs) are a component of the TME that can promote local invasion in HNSCC and other cancers. Here we report that the secretory enzyme heparan-6-O-endosulfatase 2 (Sulf-2) directly affects the CAF-supported invasion of the HNSCC cell lines SCC35 and Cal33 into Matrigel. The Sulf-2 knockout (KO) cells differ from their wild type counterparts in their spheroid growth and formation, and the Sulf-2-KO leads to decreased invasion in a spheroid co-culture model with the CAF. Next, we investigated whether a fucosylated chondroitin sulfate isolated from the sea cucumber Holothuria floridana (HfFucCS) affects the activity of the Sulf-2 enzyme. Our results show that HfFucCS not only efficiently inhibits the Sulf-2 enzymatic activity but, like the Sulf-2 knockout, inhibits Matrigel invasion of SCC35 and Cal33 cells co-cultured with primary HNSCC CAF. These findings suggest that the heparan-6-O-endosulfatases regulate local invasion and could be therapeutically targeted with the inhibitory activity of a marine glycosaminoglycan.

Keywords:  Sulf-2 CRISPRCas9 knockout; Sulf-2 inhibitor; cancer invasion; cancer-associated fibroblast; head and neck squamous cell carcinoma; heparan-6-O-endosulfatase

DOI:  https://doi.org/10.3390/cancers15215168
Steroids. 2023 Nov 09. pii: S0039-128X(23)00163-0. [Epub ahead of print] 109335

Estrogen Sulfotransferase and Sulfatase in Steroid Homeostasis, Metabolic Disease, and Cancer.

Jingyuan Wang, Ye Feng, Brian Liu, Wen Xie.

Sulfation and desulfation of steroids are opposing processes that regulate the activation, metabolism, excretion, and storage of steroids, which account for steroid homeostasis. Steroid sulfation and desulfation are catalyzed by cytosolic sulfotransferase and steroid sulfatase, respectively. By modifying and regulating steroids, cytosolic sulfotransferase (SULT) and steroid sulfatase (STS) are also involved in the pathophysiology of steroid-related diseases, such as hormonal dysregulation, metabolic disease, and cancer. The estrogen sulfotransferase (EST, or SULT1E1) is a typical member of the steroid SULTs. This review is aimed to summarize the roles of SULT1E1 and STS in steroid homeostasis and steroid-related diseases.

DOI: https://doi.org/10.1016/j.steroids.2023.109335
Front Cell Dev Biol. 2023 ;11 1271455

On-demand chlorine dioxide solution enhances odontoblast differentiation through desulfation of cell surface heparan sulfate proteoglycan and subsequent activation of canonical Wnt signaling.

Toshihiro Inubushi, Nag Priyanka, Masakatsu Watanabe, Yusuke Takahashi, Shinnosuke Kusano, Hiroshi Kurosaka, Silvana Papagerakis, Petros Papagerakis, Mikako Hayashi, Takashi Yamashiro.

  Heparan sulfate proteoglycans (HSPGs) surround the surface of odontoblasts, and their modification affects their affinity for Wnt ligands. This study proposes applying Matching Transformation System® (MA-T), a novel chlorinated oxidant, to enhance dentinogenesis. MA-T treatment in odontoblasts decreased sulfation of HSPG and upregulated the expression of dentin sialophosphoprotein (Dspp) and Dentin Matrix Protein 1 (Dmp1) via activation of canonical Wnt signaling in vitro. Ex vivo application of MA-T also enhanced dentin matrix formation in developing tooth explants. Reanalysis of a public single-cell RNA-seq dataset revealed significant Wnt activity in the odontoblast population, with enrichment for Wnt10a and Wnt6. Silencing assays showed that Wnt10a and Wnt6 were redundant in inducing Dspp and Dmp1 mRNA expression. These Wnt ligands' expression was upregulated by MA-T treatment, and TCF/LEF binding sites are present in their promoters. Furthermore, the Wnt inhibitors Notum and Dkk1 were enriched in odontoblasts, and their expression was also upregulated by MA-T treatment, together suggesting autonomous maintenance of Wnt signaling in odontoblasts. This study provides evidence that MA-T activates dentinogenesis by modifying HSPG and through subsequent activation of Wnt signaling.

Keywords:  DMP1; DSPP; MA-T; chlorinated oxidant; heparan sulfate proteoglycans

DOI:  https://doi.org/10.3389/fcell.2023.1271455
Res Sq. 2023 Oct 27. pii: rs.3.rs-3471389. [Epub ahead of print]

The role of genetic polymorphisms in the sulfation of pregnenolone by human cytosolic sulfotransferase SULT2B1a.

Eid Alatwi, Ahsan Bairam.

Pregnenolone is a key intermediate in the biosynthesis of many steroid hormones and neuroprotective steroids. Sulfotransferase family cytosolic 2B member 1 (SULT2B1a) has been reported to be highly selective to sulfate pregnenolone. This study aimed to clarify the effect of missense single nucleotide polymorphisms (SNPs) of the human SULT2B1 gene on the sulfating activity of coded SULT2B1a allozymes toward Pregnenolone. To investigate the effects of single nucleotide polymorphisms of the SULT2B1 gene on the sulfation of pregnenolone by SULT2B1a allozymes, 13 recombinant SULT2B1a allozymes were generated, expressed, and purified using established procedures. Human SULT2B1a SNPs were identified by a comprehensive database search. 13 SULT2B1a nonsynonymous missense coding SNPs (cSNPs) were selected, and site-directed mutagenesis was used to generate the corresponding cDNAs, packaged in pGEX-2TK expression vector, encoding these 13 SULT2B1a allozymes, which were bacterially expressed in BL21 E. coli cells and purified by glutathione-Sepharose affinity chromatography. Purified SULT2B1a allozymes were analyzed for sulfating activities towards pregnenolone. In comparison with the wild-type SULT2B1a, of the 13 allozymes, 11 showed reduced activity toward pregnenolone at 0.1 µM. Specifically, P134L and R259Q allozymes, reported to be involved in autosomal-recessive congenital ichthyosis, displayed low activity (1-10%) toward pregnenolone. The findings of this study may demonstrate the impact of genetic polymorphism on the sulfation of pregnenolone in individuals with different SULT2B1 genotypes.

DOI: https://doi.org/10.21203/rs.3.rs-3471389/v1
Endocr Connect. 2023 Nov 01. pii: EC-23-0250. [Epub ahead of print]

Sex steroid levels in corresponding cerebrospinal fluid and serum samples quantified by mass spectrometry in men.

Henrik Ryberg, Anna-Karin Norlén, Andreas Landin, Per M Johansson, Zeinab Salman, Anders Wallin, Johan Svensson, Claes Ohlsson.

OBJECTIVE: Sex steroids exert important biological functions within the central nervous system (CNS), but the underlying mechanisms are poorly understood. The contribution of circulating sex steroids to the levels in CNS tissue and cerebrospinal fluid (CSF) has been sparsely investigated in human and with inconclusive results. This could partly be due to lack of sensitive validated assays. To address this, we validated a gas chromatographic-mass spectrometric (GC-MS/MS) assay for quantification of sex steroid hormones/precursors in CSF.
METHODS: GC-MS/MS quantification of dihydrotestosterone (DHT, CSF lower limit of quantification, 1.5 pg/mL), testosterone (T, 4.9), estrone (E1, 0.88), estradiol (E2, 0.25), dehydroepiandrosterone (DHEA, 38.4), androstenedione (4D, 22.3) and progesterone (P, 4.2) in CSF, and corresponding serum samples from 47 men.
RESULTS: Analyses of CSF revealed that DHEA was the major sex steroid (73.5±31.7 pg/mL) followed by 4D (61.4±29.6 pg/mL) and T (49.5±18.9 pg/mL). The CSF levels of DHT, E2 and E1 were substantially lower, and P was in general not detectable in CSF. For all sex steroids except E2, strong associations between corresponding CSF and serum levels were observed. We propose that T in CSF is derived from circulating T, DHT in CSF is from local conversion from T, while E2 in CSF is from local conversion from 4D in CNS.
CONCLUSIONS: We describe the first thoroughly validated high-sensitive mass spectrometric assay for a broad sex steroid hormone panel suitable for human CSF. This assay constitutes a new tool for investigation of the role of sex steroid hormones in the human CNS.

DOI: https://doi.org/10.1530/EC-23-0250
J Biomed Res. 2023 Nov 15. 418-430

Galectin-14 promotes hepatocellular carcinoma tumor growth via enhancing heparan sulfate proteoglycan modification.

Liming Gou, Gang Yang, Sujuan Ma, Tong Ding, Luan Sun, Fang Liu, Jin Huang, Wei Gao.

  Hepatocellular carcinoma (HCC) is a highly heterogeneous malignancy and lacks effective treatment. Bulk-sequencing of different gene transcripts by comparing HCC tissues and adjacent normal tissues provides some clues for investigating the mechanisms or identifying potential targets for tumor progression. However, genes that are exclusively expressed in a subpopulation of HCC may not be enriched or detected through such a screening. In the current study, we performed a single cell-clone-based screening and identified galectin-14 as an essential molecule in the regulation of tumor growth. The aberrant expression of galectin-14 was significantly associated with a poor overall survival of liver cancer patients with database analysis. Knocking down galectin-14 inhibited the proliferation of tumor growth, whereas overexpressing galectin-14 promoted tumor growth in vivo. Non-targeted metabolomics analysis indicated that knocking down galectin-14 decreased glycometabolism; specifically that glycoside synthesis was significantly changed. Further study found that galectin-14 promoted the expression of cell surface heparan sulfate proteoglycans (HSPGs) that functioned as co-receptors, thereby increasing the responsiveness of HCC cells to growth factors, such as epidermal growth factor and transforming growth factor-alpha. In conclusion, the current study identifies a novel HCC-specific molecule galectin-14, which increases the expression of cell surface HSPGs and the uptake of growth factors to promote HCC cell proliferation.

Keywords:  co-receptor; galectin-14; heparan sulfate proteoglycans; hepatocellular carcinoma

DOI:  https://doi.org/10.7555/JBR.37.20230085
J Biomol Struct Dyn. 2023 Nov 17. 1-7

Insights into the molecular mechanism of dextran sulfate inhibiting DNA digestion by pepsin: a crucial role of sulfate group on the binding to DNA.

Jing Zhang, Leshan Ding, Hu Hou, Long Yu, Jing Li, Ping Dong.

  Sulfate polysaccharides can inhibit DNA digestion in simulated gastric juice in vitro, which is important for regulating dietary nucleic acids metabolism, but the mechanism of inhibition is unclear. This study used dextran sulfate (DS) with different sulfate groups and molecular weights to explore the effect of DS on DNA digestion. Molecular interactions between DS and DNA were investigated by biolayer interferometry (BLI), isothermal titration calorimetry (ITC) and molecular dynamics simulations. Results indicated that DS with higher molecular weight and sulfate group content showed stronger inhibitory effect of DNA digestion. ITC results showed that the combined Kd value of DNA and DS was about 2.53 mM. The main reason for inhibition of DNA digestion is that the formation of hydrogen bonds between the sulfate group of DS and DNA bases hinders the binding of DNA to pepsin. This finding will facilitate new strategies for nucleic acid metabolism and oral drug delivery.Communicated by Ramaswamy H. Sarma.

Keywords:  DNA digestion; dextran sulfate; inhibitory effect; interaction; molecular mechanism

DOI:  https://doi.org/10.1080/07391102.2023.2283145
Biomed Rep. 2023 Dec;19(6): 99

Octanoyl esterification of low molecular weight sulfated galactan enhances the cellular uptake and collagen expression in fibroblast cells.

Waraporn Sakaew, Somsuda Somintara, Kamonwan Jongsomchai, Jamal El-Abid, Kanokpan Wongprasert, José Kovensky, Tawut Rudtanatip.

  Low molecular weight sulfated galactan (LMSG) supplemented with octanoyl ester (Oct-LMSG) demonstrated superior wound healing activity compared to the unsupplemented LMSG in a fibroblast wound model. To test the hypothesis that the increased bioactivity of Oct-LMSG may depend on its penetration into the plasma membrane, its cellular uptake was investigated and collagen production in fibroblast cells was assessed for the first time. The cellular uptake of Oct-LMSG was examined using indirect immunofluorescence and a confocal laser scanning microscope. In addition, the degree of fibroblast activation associated with this uptake was evaluated. The results indicated increased LMSG internalization in fibroblasts treated with Oct-LMSG. Transmission electron micrographs revealed the ultrastructure of active protein production in fibroblasts upon treatment with Oct-LMSG. In addition, Oct-LMSG upregulated the expression of type I collagen mRNA and proteins, as well as related signaling molecules involved in collagen synthesis, including collagen type I α1 chain (Col1A1), Col1A2, phosphorylated (p)-Smad2/3 and p-Smad4. The current findings support the notion that the supplementation of LMSG with octanoyl enhanced its cellular uptake into fibroblasts and, as a result, regulated the expression of type I collagen in fibroblasts via the activation of the Smad signaling pathway. This study demonstrates the therapeutic potential of Oct-LMSG in promoting tissue regeneration.

Keywords:  Smad proteins; collagen-I; fibroblasts; low molecular weight sulfated galactan; octanoyl ester

DOI:  https://doi.org/10.3892/br.2023.1681