Archives of biochemistry and biophysics最新文献

{"title":"Vitamin C is essential for proper myogenic differentiation","authors":"Yoshitaka Kondo ,&nbsp;Ayami Sato ,&nbsp;Noritsugu Osakabe ,&nbsp;Tatsuki Minowa ,&nbsp;Yung-Li Hung ,&nbsp;Shuichi Machida ,&nbsp;Akihito Ishigami","doi":"10.1016/j.abb.2025.110704","DOIUrl":"10.1016/j.abb.2025.110704","url":null,"abstract":"<div><h3>Background</h3><div>Vitamin C (VC) is naturally present in the blood and skeletal muscles. However, conventional myoblast culture media typically lack VC. Because VC exists in skeletal muscle, VC-supplemented media should better represent physiological conditions than VC-deficient media. Therefore, we used a VC-supplemented culture medium to examine the effects of VC deficiency on myogenic differentiation.</div></div><div><h3>Methods</h3><div>Mouse C2C12 myoblasts were cultured in VC-supplemented or VC-free differentiation medium (DM), with the medium replaced every 24 h to preserve the efficacy of VC.</div></div><div><h3>Results</h3><div>First, we confirmed that VC was reliably taken up by the C2C12 cells. We assessed the expression of muscle regulatory factors during myogenic differentiation. The expression levels of late-stage differentiation markers, including myogenin (<em>MyoG</em>), myomaker (<em>Mymk</em>), myosin heavy chain 1 (<em>Myh1</em>), and <em>Myh4</em> were elevated in VC-free DM during the early stages of myogenic differentiation. In contrast, the expression levels of terminal myogenic markers in mature myofibrils, such as troponin I slow skeletal muscle (<em>Tnni1</em>) and troponin I fast skeletal muscle (<em>Tnni2</em>), increased in cells differentiated in VC-free DM but were lower than those in cells differentiated in VC-supplemented DM. The diameters of the differentiated myotubes were smaller in VC-free DM than in VC-supplemented DM. The levels of 5-hydroxymethylcytosine (5-hmC), a product of the VC-dependent DNA demethylation enzyme ten-eleven translocation (Tet), were markedly lower in VC-free DM.</div></div><div><h3>Conclusion</h3><div>These results suggest that VC modulates myogenic differentiation.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"776 ","pages":"Article 110704"},"PeriodicalIF":3.0,"publicationDate":"2025-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145800053","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Glyceraldehyde-3-phosphate dehydrogenase is inhibited by binding of Cu(I) to the essential active site cysteine","authors":"Gustavo Pelicoli Riboldi ,&nbsp;Samantha J. Firth ,&nbsp;Arnaud Baslé ,&nbsp;Kevin J. Waldron","doi":"10.1016/j.abb.2025.110707","DOIUrl":"10.1016/j.abb.2025.110707","url":null,"abstract":"<div><div>Copper is an essential micronutrient for bacteria, needed for important copper enzymes such as terminal respiratory oxidases. However, in excess, copper is toxic to bacteria. This toxicity is caused by its ability to bind tightly to proteins through the formation of Cu-Cys and Cu-His bonds. To control toxicity, bacteria have evolved homeostatic systems to safely handle the copper they need while efficiently sequestering and effluxing excess copper ions. We previously found that GapA, the abundant glycolytic glyceraldehyde-3-phosphate dehydrogenase enzyme in the <em>Staphylococcus aureus</em> cytosol, becomes associated with copper within cells cultured in medium containing excess copper. We found that this association of GapA with copper resulted in inhibition of its enzyme activity. Here, we have characterised this binding of copper ions to <em>S. aureus</em> GapA <em>in vitro</em> to determine the mechanism of copper inhibition of GapA. We found that purified recombinant GapA binds a single Cu(I) ion with high affinity. Crystallographic structural determination showed association of this copper ion with two active site residues, Cys151 and His178, known to be important for catalysis. This observation was confirmed by characterisation of mutated variants lacking these residues, which showed reduced ability to bind Cu(I) ions. Finally, we demonstrated that the cytosolic copper metallochaperone, CopZ, exhibits a tighter affinity for Cu(I) and can remove copper from GapA <em>in vitro</em>. Together, our data demonstrate the mechanism by which excess copper binds to the <em>S. aureus</em> GapA enzyme and irreversibly inhibit its activity and how the cellular homeostasis system is capable of resolving this inhibition.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"776 ","pages":"Article 110707"},"PeriodicalIF":3.0,"publicationDate":"2025-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145793148","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Quercetin enhances muscle FNDC5/irisin, partly through AMPK activation, and induces white adipose tissue browning in high-fat-fed rats","authors":"Victoria Muscia Saez ,&nbsp;Diahann Jeanette Perdicaro ,&nbsp;Cecilia Rodriguez Lanzi ,&nbsp;Eleonora Cremonini ,&nbsp;Patricia Oteiza ,&nbsp;Marcela Alejandra Vazquez Prieto","doi":"10.1016/j.abb.2025.110708","DOIUrl":"10.1016/j.abb.2025.110708","url":null,"abstract":"<div><div>Irisin is a myokine released during physical activity that contributes to the browning of white adipose tissue (WAT), providing various metabolic benefits. We previously demonstrated that supplementation with the flavonoid quercetin (Q) alleviates high-fat diet (HFD) induced visceral WAT hypertrophy, inflammation, and insulin resistance in rats. In this study, we aimed to investigate whether these beneficial effects might be associated with the ability of Q to upregulate muscle fibronectin type 3 domain-containing protein 5 (FNDC5)/irisin and uncoupling protein-1 (UCP-1), along with other brown fat markers in inguinal WAT (iWAT). HFD-fed rats for 6 weeks showed reduced muscle protein levels of FNDC5 and peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) compared to the control (Ctrl) group. Q supplementation (20 mg/kg/body weight (BW)/day) reverted these adverse effects of the HFD, significantly increasing muscle p-AMPK as well as plasma irisin levels. Furthermore, Q upregulated key proteins involved in WAT browning (PR domain containing 16 (PRDM16), PGC-1α, PPARɣ and UCP-1) and mitigated HFD-induced iWAT mass gain and hypertrophy. To further investigate the effects of Q on the FNDC5/irisin pathway, L6 myotubes were stimulated with palmitate. Q (1 μM) prevented palmitate-induced decreased levels of p-AMPK and FNDC5, and cell media secreted irisin. Moreover, Q prevented palmitate-mediated decreased Pgc1α and Fndc5 mRNA levels. Transfecting L6 myotubes with Pgc-1α siRNA revealed that Pgc-1α was involved in Q-mediated upregulation of FNDC5. Overall, results show that Q could attenuate HFD-induced iWAT hypertrophy partly through the upregulation of the FNDC5/irisin pathway in muscle and promoting WAT browning. Consumption of Q-rich foods may help mitigate adiposity associated with the consumption of diets rich in saturated fats.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"776 ","pages":"Article 110708"},"PeriodicalIF":3.0,"publicationDate":"2025-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145779789","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The SNCA-AS1/miR-138-5p/HIF1A Axis: Implications for diagnosis and cellular pathogenesis in acute cerebral infarction","authors":"Pingting Chen ,&nbsp;Yilin Feng ,&nbsp;Cu Bai","doi":"10.1016/j.abb.2025.110705","DOIUrl":"10.1016/j.abb.2025.110705","url":null,"abstract":"<div><h3>Background</h3><div>Acute cerebral infarction (ACI) is a common cerebrovascular disease with complex pathogenesis. The role of non-coding RNAs in ACI warrants further investigation. To investigate the expression, diagnostic value, and molecular mechanism of SNCA-AS1 in ACI.</div></div><div><h3>Methods</h3><div>Blood samples from 125 ACI patients and matched controls were collected. RT-qPCR detected SNCA-AS1, miR-138-5p, and HIF1A expression. ROC curves assessed diagnostic value. BV-2 cells were cultured under oxygen-glucose deprivation (OGD) conditions, with RT-qPCR used to measure molecular expression. Dual-luciferase assays and RNA immunoprecipitation (RIP) were employed to verify the intermolecular interactions within the axis. ELISA determined cytokine levels, CCK-8 and Transwell assays evaluated proliferation and migration, and flow cytometry detected apoptosis. Data were statistically analyzed using t-tests, ANOVA, and correlation analysis.</div></div><div><h3>Results</h3><div>SNCA-AS1 and HIF1A were upregulated, while miR-138-5p was downregulated in ACI patients. SNCA-AS1 showed a diagnostic AUC of 0.849. Experiments confirmed SNCA-AS1 binds miR-138-5p and regulates HIF1A. SNCA-AS1 or HIF1A knockdown alleviated inflammation, suppressed proliferation and migration, and promoted apoptosis, effects reversed by miR-138-5p inhibitor.</div></div><div><h3>Conclusion</h3><div>SNCA-AS1 participates in ACI pathogenesis through the miR-138-5p/HIF1A axis and may serve as a potential diagnostic marker.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"778 ","pages":"Article 110705"},"PeriodicalIF":3.0,"publicationDate":"2025-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145773347","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"LncRNA SNHG6 attenuates ferroptosis in high glucose-treated renal tubular epithelial cells by stabilizing YY1 to activate the PI3K/AKT/GSK-3β pathway","authors":"Qiguo Wang ,&nbsp;Qin Wang ,&nbsp;Xiangyu Meng ,&nbsp;Xiaoman Ji ,&nbsp;Ting Wang","doi":"10.1016/j.abb.2025.110702","DOIUrl":"10.1016/j.abb.2025.110702","url":null,"abstract":"<div><div>Ferroptosis has emerged as a critical mechanism contributing to renal tubular epithelial cell injury in diabetic nephropathy (DN), though its mechanisms require further elucidation. This study investigated the expression and regulatory role of long non-coding RNA small nucleolar RNA host gene 6 (SNHG6) in DN-associated ferroptosis. Bioinformatics analysis confirmed the significant downregulation of SNHG6 in DN and its diagnostic value, while real-time quantitative PCR experiments demonstrated that high glucose treatment for 24 h induced the downregulation of SNHG6 in renal tubular epithelial cells HK-2. Overexpression of SNHG6 alleviated high glucose-triggered ferroptosis of HK-2 cells by promoting the expression of glutathione peroxidase 4 and reducing the expression of acyl-CoA synthetase long chain family member 4 and transferrin receptor, manifested by increased cell viability, decreased lactate dehydrogenase activity, and suppressed lipid peroxidation. Mechanistic studies revealed that SNHG6 stabilized YY1 mRNA by promoting its binding to the FUS RNA-binding protein, subsequently activating the PI3K/AKT/GSK-3β signaling pathway to exert a protective effect. Blocking the YY1-PI3K signaling cascade abolished SNHG6 overexpression-mediated inhibition of high glucose-induced cellular ferroptosis. In summary, SNHG6 exerts renoprotective effects in DN by modulating ferroptosis through the YY1-PI3K/AKT/GSK-3β axis.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"776 ","pages":"Article 110702"},"PeriodicalIF":3.0,"publicationDate":"2025-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145740661","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring the multiple mechanisms of Hydroxytyrosol in treating obesity","authors":"Conghui Guan ,&nbsp;Ruilin Han ,&nbsp;Lijuan Liu ,&nbsp;Jinjin Liu ,&nbsp;Songbo Fu ,&nbsp;Xulei Tang","doi":"10.1016/j.abb.2025.110703","DOIUrl":"10.1016/j.abb.2025.110703","url":null,"abstract":"<div><h3>Background</h3><div>The exploration of multi-mechanism therapeutics may represent a key strategy for treating obesity and metabolic dysfunction-associated steatotic liver disease. Hydroxytyrosol (HT), a phenolic compound derived from olive oil in the Mediterranean diet, exhibits potential for obesity treatment; however, its precise mechanisms remain incompletely understood.</div></div><div><h3>Objective</h3><div>This study aimed to elucidate the therapeutic effects of HT in obesity and to uncover its underlying regulatory mechanisms.</div></div><div><h3>Methods</h3><div>An obesity model was induced in C57BL/6 mice via a high-fat diet (HFD) to evaluate the in vivo effects of HT. Assessments included glucose tolerance test (GTT), insulin tolerance test (ITT) and biochemical analyses of fasting blood glucose (FBG), fasting serum insulin (FINS), uric acid (UA), blood lipid, aspartate aminotransferase (AST), and alanine aminotransferase (ALT). Hepatic, skeletal muscle, myocardial and adipose tissue (AT) morphology were examined via H&amp;E staining. In vitro, 3T3-L1 preadipocytes were used to investigate the effects of HT on adipogenesis and thermogenic capacity. Lipid accumulation was assessed by Oil Red O and BODIPY 493/503 staining. Protein expression levels were determined via immunohistochemistry and Western blot analysis.</div></div><div><h3>Results</h3><div>HFD feeding led to increased body weight, FBG, FINS, UA, AST, ALT, and the area under the curve (AUC) for GTT and ITT. H&amp;E staining revealed hepatic vacuolization, skeletal muscle fiber hypertrophy, and myocardial disorganization in HFD-fed mice. HT treatment significantly reduced body weight, improved glucose homeostasis, lipid metabolism, and liver function, and restored normal tissue morphology of liver, skeletal muscle, myocardium, and AT. HFD upregulated the expression of PPARγ, C/EBPβ, FABP4, STING1, and NLRP3 proteins in white AT, which were markedly attenuated by HT. HT reversed HFD-induced downregulation of PGC1α and UCP1 in brown AT. In vitro experiments confirmed that HT modulates adipogenesis and thermogenic activation in preadipocyte via the STING1/NLRP3 pathways.</div></div><div><h3>Conclusion</h3><div>These findings demonstrate that HT acts as a novel anti-obesity agent by targeting the STING1/NLRP3 axis to suppress adipogenesis in adipose tissue and ameliorate obesity-related alterations in AT, liver, skeletal muscle, and myocardium. This study provides a mechanistic foundation for the potential application of HT in obesity intervention.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"776 ","pages":"Article 110703"},"PeriodicalIF":3.0,"publicationDate":"2025-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145751439","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"N-terminal KTKEGV motif lysine residues of α-Synuclein are critical for TLR2 interaction and activation","authors":"Krishna Singh Bisht,&nbsp;Manisha Kumari,&nbsp;Tushar Kanti Maiti","doi":"10.1016/j.abb.2025.110701","DOIUrl":"10.1016/j.abb.2025.110701","url":null,"abstract":"<div><div>Synucleinopathies, such as Parkinson's Disease, involve widespread intracellular protein aggregates. α-synuclein (αSyn) protein contributes majorly to these aggregates called as Lewy Bodies, which are linked to neurodegeneration. αSyn has been shown to transmit from infected to healthy neurons, thus spreading the pathology. During its transmission, αSyn is also known to activate microglia by interacting with the glial surface receptors such as Toll-Like receptors (TLRs). This activation is implicated in the production of interleukins and proinflammatory cytokines, exacerbating neuronal death through chronic neuroinflammation. However, the mechanism of αSyn and TLR interaction is not well elucidated. In this study, using biophysical methods and in the HEK-TLR2 cells, we demonstrate that αSyn N-terminal has a higher binding affinity with TLR2. This interaction is mediated through the lysine residues in the second and third KTKEGV motifs of αSyn. The lysine to alanine mutations in the N-terminal KTKGEV motifs perturb the αSyn/TLR2 interaction. We also demonstrate that the lysine residues of 2nd and 3rd KTKEGV motifs are critical for αSyn-mediated TLR2 activation. Our study demonstrates that the N-terminal KTKEGV lysine residues facilitate interaction with TLR2, thereby regulating αSyn-mediated neuroinflammation.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"776 ","pages":"Article 110701"},"PeriodicalIF":3.0,"publicationDate":"2025-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145740666","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The conference landscape of selenium research: A brief history, global structure, and stewardship","authors":"Lucy M. Kyung ,&nbsp;David G. Churchill","doi":"10.1016/j.abb.2025.110676","DOIUrl":"10.1016/j.abb.2025.110676","url":null,"abstract":"<div><div>The chemical element selenium is a main group element with some commonality to sulfur but with exquisite chemistry and biology that are still being figured out to this day. It was discovered to be a trace element essential to human health and is found in the enzyme active sites of Glutathione Peroxidases (GPX) and Thioredoxin Reductases (TXNRD). There is interest in using this element in chemistry by monitoring it in human health, as well as gauging its prevalence in the environment and agriculture. Selenium continues to warrant study from a wide perspective including chemistry, biochemistry, and its role in health and diseases. In this article, we consider the five main conference series that continue to bring selenium (and trace element) researchers together. We provide a history and clarify the five different conference series by making a graphical analogy to metro lines and their occasionally shared stations. We suggest color coding for <figure><img></figure>, <figure><img></figure>, <figure><img></figure>, <figure><img></figure>, and<figure><img></figure>. <figure><img></figure> was renamed <figure><img></figure> We list keywords and concepts such as health, environment, chemistry, and synthetic chemistry etc. The Selenium conferences are flourishing thanks to a relatively small group of dedicated practitioners who devote effort, not only on preparing “hard currency” journal manuscripts but also on event organization and stewardship. This article includes a table of all previous locations and dates spanning 55 years of meetings along with additional pertinent information. There are three tables and three figures. A call is made for continued leadership in the selenium field, and recognition is given to those who are currently dedicating themselves.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"777 ","pages":"Article 110676"},"PeriodicalIF":3.0,"publicationDate":"2025-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145740742","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"circCacna1c modulates osteogenic differentiation of BMSCs via miR-7669-3p/Ciart axis in osteoporosis","authors":"Zhipeng Chen ,&nbsp;Yan Li ,&nbsp;Bailing Chen","doi":"10.1016/j.abb.2025.110678","DOIUrl":"10.1016/j.abb.2025.110678","url":null,"abstract":"<div><div>Osteoporosis is a prevalent disorder characterized by reduced bone mass and deterioration of bone microstructure, leading to an increased risk of fractures. The osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) plays a critical role in bone formation. Circular RNAs (circRNAs) have been implicated in various diseases, including osteoporosis, by modulating gene expression. This study explores the role of circCacna1c in osteogenic differentiation and its potential as a therapeutic target for osteoporosis. CircCacna1c expression was downregulated in OVX mice and upregulated during osteogenic differentiation of BMSCs. Overexpression of circCacna1c promoted osteogenesis by increasing key osteogenic markers, such as <em>Runx2</em>, <em>Ocn</em>, and <em>Alp</em>. CircCacna1c was found to act as a molecular sponge for miR-7669-3p, which targets <em>Ciart</em>, enhancing its expression. The miR-7669-3p/<em>Ciart</em> axis played a crucial role in osteogenic differentiation, with knockdown of <em>Ciart</em> reversing the osteogenic-promoting effects of circCacna1c. In conclusions, the circCacna1c/miR-7669-3p/<em>Ciart</em> pathway is involved in regulating the osteogenic differentiation of BMSCs and may serve as a potential therapeutic target for osteoporosis. Modulating circCacna1c expression or its downstream signaling could offer novel strategies for the treatment and management of osteoporosis.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"776 ","pages":"Article 110678"},"PeriodicalIF":3.0,"publicationDate":"2025-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145740685","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Kinetic investigation on d-amino acid containing peptides and carboxypeptidase Y","authors":"Joshua I. Putman,&nbsp;Arzoo Patel,&nbsp;Maria Olds,&nbsp;Alaa Aziz,&nbsp;Haritha Asokan-Sheeja,&nbsp;He Dong,&nbsp;Kayunta L. Johnson-Winters,&nbsp;Daniel W. Armstrong","doi":"10.1016/j.abb.2025.110699","DOIUrl":"10.1016/j.abb.2025.110699","url":null,"abstract":"<div><div>Carboxypeptidases catalyze the hydrolysis of the peptide bond of C terminal amino acid residues. Carboxypeptidase Y can hydrolyze all 20 naturally-occurring amino acids at varying rates under optimal conditions. Previous data suggests that carboxypeptidase Y has more difficulty hydrolyzing <span>d</span>-amino acids from the C terminus of peptides than <span>l</span>-amino acids. This enzyme was used for amino acid sequence determination prior to modern proteomics. However, most modern proteomic methods assume that all peptides are comprised of <span>l<em>-</em></span>amino acids and do not distinguish L<span>-</span>from <span>d<em>-</em></span>amino acids within the peptide sequence. Most existing methods that allow for chiral differentiation require synthetic standards or incur racemization. Steady-state kinetic analysis was performed to elucidate differences in hydrolytic rates between peptides composed solely of <span>l</span>-amino acids at the carboxy-terminal and those incorporating <span>d</span>-amino acids. Our data suggests that the <span>l<em>-</em></span>amino acid exclusive peptides are hydrolyzed at a rate two-to-five orders of magnitude higher than <span>d</span>-amino acid containing peptides. Such differences are necessary to completely digest and eliminate <span>l<em>-</em></span>amino acid exclusive peptides while leaving the carboxy-terminal <span>d<em>-</em></span>amino acid peptides intact. The <span>d<em>-</em></span>amino acid containing peptides can then be preconcentrated to enhance detection limits to facilitate scouting for the desired <span>d<em>-</em></span>amino acid containing peptides. Interestingly, the peptide epimers bind equally well to carboxypeptidase Y on a low μM level, indicating high substrate affinity.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"776 ","pages":"Article 110699"},"PeriodicalIF":3.0,"publicationDate":"2025-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145740718","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}