Biochemistry (Moscow)最新文献

{"title":"Features of Photosynthesis in Arabidopsis thaliana Plants with Knocked out Genes Encoding Chloroplast Carbonic Anhydrases αCA1 and βCA1","authors":"Natalia N. Rudenko,&nbsp;Maria Yu. Ruppert,&nbsp;Lyudmila K. Ignatova,&nbsp;Elena M. Nadeeva,&nbsp;Daria V. Vetoshkina,&nbsp;Boris N. Ivanov","doi":"10.1134/S0006297925600954","DOIUrl":"10.1134/S0006297925600954","url":null,"abstract":"<p>The knockout of either <i>At3g01500</i> or <i>At3g52720</i> gene encoding <i>Arabidopsis thaliana</i> βCA1 and αCA1 carbonic anhydrase (CA), respectively, led to a lower CA activity of the chloroplast stroma preparations from the knockout mutant plants (αCA1-KO and βCA1-KO) compared to such preparations from the wild-type (WT) plants. To identify the differences in the photosynthetic characteristics of mutant and WT plants, they were grown in low light (LL; 50-70 µmol quanta∙m⁻²∙s⁻¹, natural conditions) and high light (HL; 400 µmol quanta∙m⁻²∙s⁻¹, stressful conditions). The rate of CO₂ assimilation measured at 400 µmol quanta∙m⁻²∙s⁻¹ in plants grown under LL was lower in αCA1-KO and βCA1-KO mutants compared to WT plants. The rate of photosynthetic electron transport was lower in αCA1-KO plants and higher in βCA1-KO plants than in WT plants; the content of CO₂ in chloroplasts was lower in βCA1-KO plants than in both WT and αCA1-KO plants, where it differed little. The value of the proton-motive force was higher in βCA1-KO plants and lower in αCA1-KO plants than in WT plants due to changes in ΔpH value. The obtained results suggest that βCA1 facilitates the intake of inorganic carbon into chloroplasts, while αCA1 ensures the conversion of bicarbonate into CO₂ in the chloroplast stroma for its use in the reaction catalyzed by Ribulose bisphosphate carboxylase/oxygenase (RuBisCO). In both αCA1-KO and βCA1-KO mutants, the expression levels of genes encoding other chloroplast CAs differed markedly from those in WT plants; the pattern of the changes in the genes expression depended on the light intensity during cultivation. The content of hydrogen peroxide in the leaves of both αCA1-KO and βCA1-KO mutants was higher in LL and lower in HL than in WT plants. The expression levels of stress marker genes changed similarly in both types of mutant plants. A possible involvement of the chloroplast stroma CAs in the transmission of stress signals in higher plants is discussed.</p>","PeriodicalId":483,"journal":{"name":"Biochemistry (Moscow)","volume":"90 )\nGuest","pages":"894 - 910"},"PeriodicalIF":2.2,"publicationDate":"2025-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144783155","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Photosynthetic Control and Its Role in Protection of Photosystem I against Photoinhibition","authors":"Daria V. Vilyanen,&nbsp;Marina A. Kozuleva","doi":"10.1134/S0006297925601121","DOIUrl":"10.1134/S0006297925601121","url":null,"abstract":"<p>This review addresses photosynthetic control as a protective mechanism that prevents photoinhibition of photosystem I under conditions of imbalance between CO₂ assimilation during the Calvin–Benson–Bassham cycle and light reactions in the thylakoid photosynthetic apparatus. We discuss the pathways of photosystem I photoinhibition and describe protective mechanisms that prevent photodamage of photosystem I. We propose a hypothesis regarding the influence of photosynthetic control on formation of reactive oxygen species in photosystem I. pH-sensitivity of plastoquinol oxidation at the quinol-oxidizing (Qo) site of the cytochrome <i>b</i>₆<i>f</i> complex is analyzed, and function of two proton-conducting channels that release protons into the thylakoid lumen from the cytochrome <i>b</i>₆<i>f</i> complex is described. We examine impact of photosynthetic control on the functioning of the cytochrome <i>b</i>₆<i>f</i> complex itself, and propose a hypothesis regarding the preferential activation of photosynthetic control in the thylakoid grana, which ensures operation of the cyclic electron transport around photosystem I as a main protective mechanism.</p>","PeriodicalId":483,"journal":{"name":"Biochemistry (Moscow)","volume":"90 )\nGuest","pages":"840 - 859"},"PeriodicalIF":2.2,"publicationDate":"2025-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144783157","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Short-Term Photoinhibition Induces Long-Term Hydrogen Photoproduction in a Phototrophic Culture of Chlorella sorokiniana on Complete Medium","authors":"Alena A. Volgusheva,&nbsp;Taras K. Antal","doi":"10.1134/S000629792560098X","DOIUrl":"10.1134/S000629792560098X","url":null,"abstract":"<p>This work demonstrates, for the first time, capacity of the <i>Chlorella sorokiniana</i> immobilized in alginate to produce hydrogen (H₂) over an extended period of time when cultivated under strictly photoautotropic conditions on complete mineral medium. In order to reduce photosynthetic activity, immobilized cells were subjected to a 30-minute pre-incubation period at high light intensity of 1000 μmol photons m⁻²∙s⁻¹. The ability to produce H₂ was evaluated under illumination of 40 μmol/(m²∙s). The culture not bubbled with argon produced H₂ for 9 days; total gas yield was 0.1 mol H₂/m². In the culture under argon atmosphere, the release of H₂ continued for 51 days, resulting in a total yield of 0.55 mol H₂/m². The immobilized culture was capable of H₂ production at 16% O₂ in the gas phase, which may be due to the effects of photoinhibition and activation of oxygen uptake pathways in mitochondria and chloroplast. Analysis of the functioning of electron-transport chain in the microalgae cells revealed decrease in the rate of electron transport, increase in the size of the PSII antenna, and development of non-photochemical quenching processes, while activity of PSII remained moderately high (Fv/Fm = 0.4-0.6). Inhibitor analysis using 10⁻⁵ M DCMU demonstrated that contribution of PSII to hydrogenase reaction increased from 30% on the first day of the experiment to 50% by the fourth day. Addition of 10⁻⁵ M DBMIB led to the 90% reduction in the rate of H₂ formation on both day 1 and day 4.</p>","PeriodicalId":483,"journal":{"name":"Biochemistry (Moscow)","volume":"90 )\nGuest","pages":"934 - 942"},"PeriodicalIF":2.2,"publicationDate":"2025-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144783072","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An Attempt to Increase Thermostability of the Mutant Photosynthetic Reaction Center of Cereibacter sphaeroides Using Disulfide Bonds","authors":"Tatiana Yu. Fufina,&nbsp;Lyudmila G. Vasilieva","doi":"10.1134/S0006297925600978","DOIUrl":"10.1134/S0006297925600978","url":null,"abstract":"<p>Methods of site-directed mutagenesis are successfully used in structural and functional studies of photosynthetic reaction centers (RCs). It has been noted that many mutations near electron transfer cofactors reduce temperature stability of the <i>Cereibacter sphaeroides</i> RCs and affect amount of RCs in the membranes. We previously reported [Selikhanov et al. (2023) <i>Membranes</i>, <b>25</b>, 154] that introduction of inter-subunit disulfide bridges on the periplasmic or cytoplasmic surface of the complex promotes increase in thermal stability of the <i>C. sphaeroides</i> RCs. In this work, an attempt was made to increase thermal stability of the mutant RC with the Ile M206 – Gln substitution by introducing inter-subunit disulfide bonds. This RC is of considerable interest for studying mechanisms of early electron transfer processes in RCs. The effect of mutations on the amount of RCs in chromatophores was analyzed and it was found that the I(M206)Q mutation leads to twofold decrease in the RC content in chromatophores, introduction of disulfide bonds on the cytoplasmic or periplasmic sides of the complex reduces the amount of RCs in membranes by one third, the triple substitution I(M206)Q/G(M19)C/T(L214)C reduces the amount of RCs in membranes almost 4-fold, and the substitutions I(M206)Q/V(M84)C/G(L278)C lead to disruption of RC assembly in the membrane. It was shown that introduction of the inter-subunit S-S bond on the cytoplasmic surface of the complex did not have a significant effect on thermal stability of the I(M206)Q RC. Our own and literature data on the factors influencing assembly processes and ensuring stability of the structure of integral membrane complexes are discussed.</p>","PeriodicalId":483,"journal":{"name":"Biochemistry (Moscow)","volume":"90 )\nGuest","pages":"873 - 881"},"PeriodicalIF":2.2,"publicationDate":"2025-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144783152","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cationic Antiseptics Disrupt the Functioning of the Electron-Transport Chain at the Acceptor Side in the Photosynthetic Reaction Centres of the Purple Bacterium Cereibacter sphaeroides","authors":"Evgenii P. Lukashev,&nbsp;Mahir D. Mamedov,&nbsp;Liya A. Vitukhnovskaya,&nbsp;Aida M. Mamedova,&nbsp;Peter P. Knox,&nbsp;Vladimir Z. Paschenko","doi":"10.1134/S0006297925600723","DOIUrl":"10.1134/S0006297925600723","url":null,"abstract":"<p>Using electrometric technique, the cationic antiseptic octenidine was revealed to reduce generation of transmembrane electrical potential difference in the chromatophores of photosynthetic bacterium <i>Cereibacter sphaeroides</i>. This is also confirmed by measurements of electrochromic shifts of carotenoid absorption bands in chromatophores. In reaction centers (RCs), isolated from chromatophores in the absence of external electron donors and acceptors, the rate of recombination between photooxidized bacteriochlorophyll P₈₇₀ and reduced secondary quinone acceptor Q_B, as measured by absorption changes in the near infrared region, was very weakly dependent on the presence of antiseptics, in contrast to the kinetics in the 400-600 nm spectral range, where absorption changes associated with the oxidation of P₈₇₀ and the formation of semiquinone radicals Q_A⁻ and Q_B⁻, as well as electrochromic shifts of the carotenoid and bacteriopheophytin RC absorption bands, were observed. The addition of cationic antiseptics modified the flash-induced absorbance changes in this region with the formation time of ~100-200 ms and a decay time of ~3 s. In the series: picloxydine – chlorhexidine – octenidine – miramistin, the last one was the most effective. The maximum amplitude of such changes was observed in the absorption region of the semiquinone radical around 460 nm. When electron transfer from Q_A⁻ to Q_B was blocked by o-phenanthroline, the effect disappeared. Cationic antiseptics are suggested to stimulate protonation of Q_B⁻ with the formation of a neutral Q_B⁻H⁺ complex.</p>","PeriodicalId":483,"journal":{"name":"Biochemistry (Moscow)","volume":"90 )\nGuest","pages":"882 - 893"},"PeriodicalIF":2.2,"publicationDate":"2025-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144783153","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of Cultivation Conditions on the Expression of the Exiguobacterium sibiricum Proteorhodopsin Gene","authors":"Lada E. Petrovskaya,&nbsp;Elena V. Spirina,&nbsp;Artemiy Yu. Sukhanov,&nbsp;Elena A. Kryukova,&nbsp;Evgeniy P. Lukashev,&nbsp;Rustam H. Ziganshin,&nbsp;Elizaveta M. Rivkina,&nbsp;Dmitrii A. Dolgikh,&nbsp;Mikhail P. Kirpichnikov","doi":"10.1134/S0006297925600917","DOIUrl":"10.1134/S0006297925600917","url":null,"abstract":"<p>Recombinant proteorhodopsin ESR of the gram-positive bacterium <i>Exiguobacterium sibiricum</i> isolated from permafrost deposits in northeastern Siberia binds retinal and acts as a light-dependent proton pump, but not much is known about its expression under natural conditions. In this work, expression of the <i>esr</i> gene in <i>E. sibiricum</i> cultures grown under various conditions was studied by quantitative PCR. It has been discovered that cultivation on poor media at low temperatures contributes to a significant increase in the content of the corresponding mRNA. The data obtained are confirmed by the results of the analysis of the membrane fraction of cells using label-free quantitative chromatography-mass spectrometry. Also, at 10°C, increased content of phytoene desaturases, which are involved in the biosynthesis of carotenoids, is observed. However, we were unable to detect the presence of a functional retinal-containing protein in the cells, presumably due to the lack of an enzymatic retinal synthesis system in<i> E. sibiricum</i>. The possible functions of ESR in <i>E. sibiricum</i> cells are discussed in connection with the characteristics of the extreme habitat of the bacterium. The results of this study contribute to expanding the understanding of the molecular mechanisms of microbial adaptation to environmental conditions and the potential role of microbial rhodopsins in these processes.</p>","PeriodicalId":483,"journal":{"name":"Biochemistry (Moscow)","volume":"90 )\nGuest","pages":"911 - 920"},"PeriodicalIF":2.2,"publicationDate":"2025-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144783154","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Protective Effect of α-Carbonic Anhydrase CAH3 Against Photoinhibition and Thermal Inactivation of Photosystem II in Membrane Preparations as Compared with α-Carbonic Anhydrase CA4","authors":"Vasily V. Terentyev,&nbsp;Liubov I. Trubitsina,&nbsp;Tatyana P. Khoroshaeva,&nbsp;Ivan V. Trubitsin","doi":"10.1134/S0006297925601133","DOIUrl":"10.1134/S0006297925601133","url":null,"abstract":"<p>Photosystem II (PSII) is one of the most vulnerable components of photosynthetic apparatus of the thylakoid membrane to the action of inhibitory factors. The donor side of PSII exhibits high sensitivity to photoinhibition and thermal inactivation, which leads to the loss of O₂-evolving function of the water-oxidizing complex (WOC). The data obtained in this study demonstrated increased stability of WOC activity in the PSII membrane preparations from the wild-type (WT) <i>Chlamydomonas reinhardtii</i> compared to the PSII preparations from the <i>cia3</i> mutant, which lack α-carbonic anhydrase (CA) CAH3, under conditions of moderate photoinhibition and thermal inactivation. This effect was completely eliminated by adding a CA inhibitor to the PSII preparations from WT. At the same time, addition of active recombinant CAH3 (rCAH3) protein to the preparations from <i>cia3</i> restored increased resistance of PSII to these factors. Under the same conditions of photoinhibition and thermal inactivation, the PSII preparations from <i>Arabidopsis thaliana</i> demonstrated very low loss of O₂-evolving activity, regardless of the presence or absence of carbonic anhydrase α-CA4, which is similar to CAH3. More pronounced suppression of the O₂-evolving activity in the PSII from <i>A. thaliana</i> mutants lacking CA4 was observed only when they were incubated at elevated temperature, indicating the possibility of more significant conformational changes in the WOC proteins of PSII. Despite the clear binding of the rCAH3 to PSII membrane preparations from <i>A. thaliana</i>, the enzyme had little effect on the WOC activity in these preparations, suggesting absence of functional interaction between the rCAH3 and PSII from <i>A. thaliana</i>. The obtained results indicate different mechanisms of involvement of CAH3 and CA4, both of which are assumed to exist in close association with PSII in live systems, in the PSII functioning.</p>","PeriodicalId":483,"journal":{"name":"Biochemistry (Moscow)","volume":"90 )\nGuest","pages":"860 - 872"},"PeriodicalIF":2.2,"publicationDate":"2025-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144783159","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Preface to the Special Issue “Genetic Technologies in Medicine, Agriculture, and Biotechnology”","authors":"Dmitry S. Kostyushev,&nbsp;Andrey A. Zamyatnin Jr.","doi":"10.1134/S0006297925601741","DOIUrl":"10.1134/S0006297925601741","url":null,"abstract":"","PeriodicalId":483,"journal":{"name":"Biochemistry (Moscow)","volume":"90 ).","pages":"649 - 649"},"PeriodicalIF":2.2,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144558654","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Designing a Thermostable Mini-Intein for Intein-Mediated Purification of Recombinant Proteins and Peptides","authors":"Andrey A. Karanov,&nbsp;Evgeniy A. Zayats,&nbsp;Maria A. Kostromina,&nbsp;Yulia A. Abramchik,&nbsp;Aleksandra R. Sharafutdinova,&nbsp;Maria S. Surkova,&nbsp;Andrey A. Zamyatnin Jr.,&nbsp;Roman S. Esipov","doi":"10.1134/S0006297925600358","DOIUrl":"10.1134/S0006297925600358","url":null,"abstract":"<p>This paper reports the design of a thermostable temperature-activated mini-intein based on the full-length intein DnaE1 from <i>Thermus thermophilus</i> HB27 (<i>Tth</i>DnaE1). We performed rational design of three mini-inteins <i>Tth</i>DnaE1 Δ272, Δ280, and Δ287 through deletion mutations in the full-length intein sequence. Two mini-inteins (Δ272 and Δ280) were capable of efficient protein splicing at temperatures above 50°C. The most active mini-intein with the Δ280 deletion was selected as a platform for further design of a self-cleaving carrier of affinity tags through single-point mutagenesis. Three mutations – C1A, D405G, and the combined C1A/D405G – were introduced to inhibit N-terminal extein cleavage and extein ligation. As a result, the mini-intein Δ280 with double mutation C1A/D405G displayed the highest efficiency of C-terminal extein cleavage with temperature optimum around 60°C. Thus, we constructed thermostable temperature-activated mini-intein capable of efficient protein splicing or cleavage of the C-terminal extein. The engineered <i>Tth</i>DnaE1 Δ280 C1A/D405G mini-intein can serve as a basis for the development of new expression system for intein-mediated production of pharmaceutically relevant recombinant proteins and peptides.</p>","PeriodicalId":483,"journal":{"name":"Biochemistry (Moscow)","volume":"90 ).","pages":"818 - 827"},"PeriodicalIF":2.2,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144558651","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Selection of UTRs in mRNA-Based Gene Therapy and Vaccines","authors":"Ilya A. Volkhin,&nbsp;Anastasia Iu. Paremskaia,&nbsp;Maria A. Dashian,&nbsp;Darya S. Smeshnova,&nbsp;Roman E. Pavlov,&nbsp;Olga N. Mityaeva,&nbsp;Pavel Yu. Volchkov,&nbsp;Andrei A. Deviatkin","doi":"10.1134/S0006297924604659","DOIUrl":"10.1134/S0006297924604659","url":null,"abstract":"<p>The untranslated regions (UTRs) of messenger RNAs (mRNAs) play a crucial role in regulating translational efficiency, stability, and tissue-specific expression. The review describes various applications and challenges of UTR design in the development of gene therapy and mRNA-based therapeutics. UTRs affect critical biological functions, such as mRNA stability, modulation of protein synthesis, and attenuation of immune response. Incorporating tissue-specific microRNA (miRNA)-binding sites into 3′ UTRs might improve precise targeting of transgene expression and minimize off-target effects. Nucleotide modifications (pseudouridine, N1-methyladenosine, and N4-acetylcytidine) in mRNA and UTRs in particular, improve mRNA stability and translational efficiency. At the same time, several challenges remain, such as lack of consensus on UTRs best suited for certain biomedical applications. Current efforts are focused on integrating high-throughput screening, computational modeling, and experimental validation to refine UTR-based therapeutic strategies. The review presents current information on the design of UTRs and their role in therapeutic applications, with special focus on the possibilities and limitations of existing approaches.</p>","PeriodicalId":483,"journal":{"name":"Biochemistry (Moscow)","volume":"90 ).","pages":"725 - 753"},"PeriodicalIF":2.2,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144558657","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}