Journal of biochemistry最新文献

A refined method for high-purity isolation of uterine glandular epithelial cells in mouse. 一种高纯度分离小鼠子宫腺上皮细胞的改进方法。

IF 2.1 4区生物学

Journal of biochemistry Pub Date : 2025-01-22 DOI: 10.1093/jb/mvaf006

Asmaa M Youssef, Ahmed M Moustafa, Motoharu Hamada, Mayumi Sugiura-Ogasawara, Hisashi Oishi

{"title":"A refined method for high-purity isolation of uterine glandular epithelial cells in mouse.","authors":"Asmaa M Youssef, Ahmed M Moustafa, Motoharu Hamada, Mayumi Sugiura-Ogasawara, Hisashi Oishi","doi":"10.1093/jb/mvaf006","DOIUrl":"https://doi.org/10.1093/jb/mvaf006","url":null,"abstract":"The uterine endometrium consists of luminal epithelium, glandular epithelium, and stromal cells, with uterine glands playing a pivotal role in pregnancy success among mammals. Uterine glands secrete essential factors that regulate embryo development and implantation; however, their cellular biology remains poorly understood. This study presents a refined method for isolating three distinct endometrial cell types with high purity, with a specific emphasis on glandular epithelial cells. The method combines mechanical dissociation, enzymatic digestion, and immunomagnetic separation. The isolated glandular epithelial cells were maintained in culture and exhibited proliferation in response to steroid hormones. Furthermore, estrogen responsiveness was abrogated by Estrogen Receptor 1 (Esr1) knockdown mediated by siRNA. Here, we present an efficient and reproducible method for isolating uterine glandular epithelial cells with high purity, enabling their in vitro maintenance, hormone responsiveness assessment, and functional gene knockdown. These findings establish a robust platform for advancing our understanding of uterine gland biology, facilitating detailed investigations into molecular mechanisms underlying glandular function and their critical roles in establishing pregnancy success. Future research could explore the contribution of these isolated cells to endometrial receptivity and embryo implantation.","PeriodicalId":15234,"journal":{"name":"Journal of biochemistry","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143005237","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}

引用次数: 0

High GRWD1 expression may predict clinically aggressive lower grade glioma, skin cutaneous melanoma, and kidney renal clear cell carcinoma carrying wild-type p53: a systematic study based on TCGA data analysis. GRWD1高表达可预测携带野生型p53的临床侵袭性低级别胶质瘤、皮肤黑色素瘤和肾透明细胞癌：一项基于TCGA数据分析的系统研究。

IF 2.1 4区生物学

Journal of biochemistry Pub Date : 2025-01-15 DOI: 10.1093/jb/mvaf004

Kota Kayama, Akihiro Ooga, Kouji Hasetsu, Ryoma Kokubo, Nozomi Sugimoto, Masatoshi Fujita

{"title":"High GRWD1 expression may predict clinically aggressive lower grade glioma, skin cutaneous melanoma, and kidney renal clear cell carcinoma carrying wild-type p53: a systematic study based on TCGA data analysis.","authors":"Kota Kayama, Akihiro Ooga, Kouji Hasetsu, Ryoma Kokubo, Nozomi Sugimoto, Masatoshi Fujita","doi":"10.1093/jb/mvaf004","DOIUrl":"https://doi.org/10.1093/jb/mvaf004","url":null,"abstract":"Glutamate-rich WD40 repeat containing 1 (GRWD1) is a novel oncogene/oncoprotein that downregulates the p53 tumor suppressor protein through several mechanisms. One important mechanism involves binding of GRWD1 to RPL11, which competitively inhibits the RPL11-MDM2 interaction and releases RPL11-mediated suppression of MDM2 ubiquitin ligase activity toward p53. Here, we mined the TCGA (The Cancer Genome Atlas) database to gain in-depth insight into the clinical relevance of GRWD1. We found that high expression of GRWD1 is associated with a poor prognosis for lower grade glioma (LGG) of the brain, skin cutaneous melanoma (SKCM), and kidney renal clear cell carcinoma (KIRC) carrying wild-type p53. Further investigations revealed that copy number alterations in the GRWD1 gene are one determinant of GRWD1 expression level. By contrast, even in patients with a diploid GRWD1 gene, high GRWD1 expression was associated with a poor prognosis for LGG, SKCM, and KIRC carrying wild-type p53. Additional studies suggest that some transcriptional factors may be involved in regulation of GRWD1 in cancers with a diploid GRWD1 gene. Taken together, the data presented herein suggest that high expression of GRWD1 may contribute to malignant behavior, and predict a clinically unfavorable prognosis for LGG, SKCM, and KIRC carrying wild-type p53.","PeriodicalId":15234,"journal":{"name":"Journal of biochemistry","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142983613","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}

引用次数: 0

Identification of a novel Eps 15 homology domain-containing protein 1 (EHD1) and EHD4-binding motif in phostensin. 光导素中新的EHD1和ehd4结合基序的鉴定。

IF 2.1 4区生物学

Journal of biochemistry Pub Date : 2025-01-08 DOI: 10.1093/jb/mvaf002

Kuang-Yung Huang, Hui-Chun Yu, Ming-Chi Lu, Hsien-Yu Huang Tseng, Jyun-Jie Shen, Chia-Ying Lin, Pin-Chen Chen, Ya-Ting Shen, Pei-Rong Chung, Hsiao-Kuei Tsai, Si-Ru Zhou, Chia-Lin Wang, Ning-Sheng Lai, Ta-Hsien Lin, Hsien-Bin Huang

{"title":"Identification of a novel Eps 15 homology domain-containing protein 1 (EHD1) and EHD4-binding motif in phostensin.","authors":"Kuang-Yung Huang, Hui-Chun Yu, Ming-Chi Lu, Hsien-Yu Huang Tseng, Jyun-Jie Shen, Chia-Ying Lin, Pin-Chen Chen, Ya-Ting Shen, Pei-Rong Chung, Hsiao-Kuei Tsai, Si-Ru Zhou, Chia-Lin Wang, Ning-Sheng Lai, Ta-Hsien Lin, Hsien-Bin Huang","doi":"10.1093/jb/mvaf002","DOIUrl":"https://doi.org/10.1093/jb/mvaf002","url":null,"abstract":"Phostensin (PTS) encoded by KIAA1949 binds to protein phosphatase 1, F-actin, Eps 15 homology domain-containing protein 1 (EHD1) and EHD4. Most EHD-binding proteins contain a consensus motif, Asn-Pro-Phe (NPF), which interacts with the C-terminal EH domain of EHD proteins. Nevertheless, the NPF motif is absent in PTS. The binding motif for PTS to interact with EHD1 (or EHD4) remained unknown. Here, we identified that PTS-α binds to EHD1 (or EHD4) through the region of residues 51-80 which contains a consensus motif, 64ILV(X)4(L/V)RL74S. This novel consensus motif is also found in vacuolar protein sorting-35 (vps35). Replacement of 64ILV(X)4(L/V)RL74S with 64AAA(X)4(L/V)RL74S or with 64ILV(X)4AEA74A significantly reduces the binding efficiency of PTS-α to either EHD1 or EHD4 in GST pull-down assay and far western blotting assay. In addition, replacement of 218ILV(X)4VRL228S with 218AAA(X)4AEA228A decreases the binding ability of vps35 to EHD4 in far western blotting assay. Overexpression of the PTS-β in 293T cells attenuated the endocytic trafficking of transferrin. However, this attenuation of transferrin in endocytic trafficking was disrupted when 293T cells overexpressed the mutant PTS-β with a defective EHD-binding motif, suggesting that PTS-β can regulate the endocytic recycling via associating with EHD1 or EHD4.","PeriodicalId":15234,"journal":{"name":"Journal of biochemistry","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2025-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142949467","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}

引用次数: 0

SSB promotes DnaB helicase passage through DnaA complexes at the replication origin oriC for bidirectional replication. SSB促进dna解旋酶通过复制起点处的dna复合体进行双向复制。

IF 2.1 4区生物学

Journal of biochemistry Pub Date : 2025-01-08 DOI: 10.1093/jb/mvaf003

Yusuke Akama, Ryusei Yoshida, Shogo Ozaki, Hironori Kawakami, Tsutomu Katayama

{"title":"SSB promotes DnaB helicase passage through DnaA complexes at the replication origin oriC for bidirectional replication.","authors":"Yusuke Akama, Ryusei Yoshida, Shogo Ozaki, Hironori Kawakami, Tsutomu Katayama","doi":"10.1093/jb/mvaf003","DOIUrl":"https://doi.org/10.1093/jb/mvaf003","url":null,"abstract":"For bidirectional replication in E. coli, higher-order complexes are formed at the replication origin oriC by the initiator protein DnaA, which locally unwinds the left edge of oriC to promote the loading of two molecules of DnaB onto the unwound region via dynamic interactions with the helicase-loader DnaC and the oriC-bound DnaA complex. One of the two helicases must translocate rightwards through oriC-bound DnaA complex. Here, we used a synthetic forked oriC DNA, which mimics the unwound state of oriC, to examine DnaB translocation through the oriC-bound DnaA complex. We found that DnaB helicase alone cannot pass through the oriC-bound DnaA complex without the help of single strand-binding protein (SSB). In the presence of SSB, DnaB passed through this complex along with its helicase function, releasing DnaA molecules. In addition, DnaB helicase activity is known to be inhibited by oversupply of DnaC, but this inhibition was relieved by SSB. These results suggest a mechanism that when two DnaB helicases are loaded at oriC, one translocates leftwards to expand the DnaA-unwound region and allows SSB binding to the single-stranded DNA, and such SSB molecules then stimulate translocation of the other helicase rightwards through the oriC-bound DnaA complex.","PeriodicalId":15234,"journal":{"name":"Journal of biochemistry","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2025-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142949469","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}

引用次数: 0

Functional Diversity of Senescent Cells in Driving Aging Phenotypes and Facilitating Tissue Regeneration. 衰老细胞在驱动衰老表型和促进组织再生中的功能多样性。

IF 2.1 4区生物学

Journal of biochemistry Pub Date : 2025-01-06 DOI: 10.1093/jb/mvae098

Yasuhiro Nakano, Yoshikazu Johmura

{"title":"Functional Diversity of Senescent Cells in Driving Aging Phenotypes and Facilitating Tissue Regeneration.","authors":"Yasuhiro Nakano, Yoshikazu Johmura","doi":"10.1093/jb/mvae098","DOIUrl":"https://doi.org/10.1093/jb/mvae098","url":null,"abstract":"As the global population continues to age, understanding the complex role of cellular senescence and its implications in healthy lifespans has gained increasing prominence. Cellular senescence is defined as the irreversible cessation of cell proliferation, accompanied by the secretion of a range of pro-inflammatory factors, collectively termed the senescence-associated secretory phenotype (SASP), in response to various cellular stresses. While the accumulation of senescent cells has been strongly implicated in the aging process and the pathogenesis of age-related diseases owing to their pro-inflammatory properties, recent research has also highlighted their essential roles in processes such as tumour suppression, tissue development, and repair. This review provides a comprehensive examination of the dual nature of senescent cells, evaluating their deleterious contributions to chronic inflammation, tissue dysfunction, and disease, as well as their beneficial roles in maintaining physiological homeostasis. Additionally, we explored the therapeutic potential of senolytic agents designed to selectively eliminate detrimental senescent cells while considering the delicate balance between transient and beneficial senescence and the persistence of pathological senescence. A deeper understanding of these dynamics is critical to develop novel interventions aimed at mitigating age-related dysfunctions and enhancing healthy life expectancies.","PeriodicalId":15234,"journal":{"name":"Journal of biochemistry","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2025-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142931763","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}

引用次数: 0

Exploring the roles of Lem2 and Bqt4 in lipid metabolism for nuclear envelope maintenance: a novel perspective. 探索 Lem2 和 Bqt4 在维持核包膜的脂质代谢中的作用：一个新的视角

IF 2.1 4区生物学

Journal of biochemistry Pub Date : 2025-01-06 DOI: 10.1093/jb/mvae072

Kei-Ichiro Ishiguro

引用次数: 0

Cellular Senescence in The Cancer Microenvironment. 癌症微环境中的细胞衰老。

IF 2.1 4区生物学

Journal of biochemistry Pub Date : 2025-01-06 DOI: 10.1093/jb/mvaf001

Satoru Meguro, Makoto Nakanishi

引用次数: 0

Open and closed structures of L-arginine oxidase by cryo-electron microscopy and X-ray crystallography. 通过冷冻电镜和 X 射线晶体学研究 L-精氨酸氧化酶的开放和封闭结构。

IF 2.1 4区生物学

Journal of biochemistry Pub Date : 2025-01-06 DOI: 10.1093/jb/mvae070

Hiroki Yamaguchi, Kazutoshi Takahashi, Nobutaka Numoto, Hiroshi Suzuki, Moemi Tatsumi, Akiko Kamegawa, Kouki Nishikawa, Yasuhisa Asano, Toshimi Mizukoshi, Hiroshi Miyano, Yoshinori Fujiyoshi, Masayuki Sugiki

{"title":"Open and closed structures of L-arginine oxidase by cryo-electron microscopy and X-ray crystallography.","authors":"Hiroki Yamaguchi, Kazutoshi Takahashi, Nobutaka Numoto, Hiroshi Suzuki, Moemi Tatsumi, Akiko Kamegawa, Kouki Nishikawa, Yasuhisa Asano, Toshimi Mizukoshi, Hiroshi Miyano, Yoshinori Fujiyoshi, Masayuki Sugiki","doi":"10.1093/jb/mvae070","DOIUrl":"10.1093/jb/mvae070","url":null,"abstract":"L-arginine oxidase (AROD, EC 1.4.3.25) is an oxidoreductase that catalyses the deamination of L-arginine, with flavin adenine dinucleotide (FAD) as a cofactor. Recently identified AROD from Pseudomonas sp. TPU 7192 (PT-AROD) demonstrates high selectivity for L-arginine. This enzyme is useful for accurate assays of L-arginine in biological samples. The structural characteristics of the FAD-dependent AROD, however, remain unknown. Here, we report the structure of PT-AROD at a resolution of 2.3 Å by cryo-electron microscopy. PT-AROD adopts an octameric structure with D4 symmetry, which is consistent with its molecular weight in solution, estimated by mass photometry. Comparative analysis of this structure with that determined using X-ray crystallography reveals open and closed forms of the lid-like loop at the entrance to the substrate pocket. Furthermore, mutation of Glu493, located at the substrate binding site, diminishes substrate selectivity, suggesting that this residue contributes significantly to the high selectivity of PT-AROD.","PeriodicalId":15234,"journal":{"name":"Journal of biochemistry","volume":" ","pages":"27-36"},"PeriodicalIF":2.1,"publicationDate":"2025-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11694665/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142466392","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Production of non-triple helical collagen polypeptides under Hypoxia and the Implication for Tumor. 缺氧条件下非三螺旋胶原多肽的生成及其对肿瘤的意义。

IF 2.1 4区生物学

Journal of biochemistry Pub Date : 2025-01-04 DOI: 10.1093/jb/mvae099

Kosuke Sekine, Kazuhiro Tokunaka, Arihiro Tomura, Hidemitsu Sugihara, Yuki Saijo, Yongchol Shin, Toshihiko Hayashi, Makoto Morita, Yasutada Imamura

{"title":"Production of non-triple helical collagen polypeptides under Hypoxia and the Implication for Tumor.","authors":"Kosuke Sekine, Kazuhiro Tokunaka, Arihiro Tomura, Hidemitsu Sugihara, Yuki Saijo, Yongchol Shin, Toshihiko Hayashi, Makoto Morita, Yasutada Imamura","doi":"10.1093/jb/mvae099","DOIUrl":"https://doi.org/10.1093/jb/mvae099","url":null,"abstract":"Non-triple helical collagen polypetides (NTHs) are alternative gene products lacking the typical collagen triple-helical structure. This study investigated NTH production in tumor cells and tissues. NTH α1(IV) was detected in various human tumor cell lines and extracted from human lung cancer tissues and tumors in mice. NTH production was significantly affected by serum concentration and occurred under hypoxic or hypoxia-mimetic conditions, even with sufficient ascorbic acid. This suggests NTHs are produced under physiological hypoxia, potentially contributing to tumor angiogenesis. NTH production generally coincided with hypoxia-inducible factor-1α (HIF-1α) accumulation, except with cobalt chloride, indicating HIF-1α isn't directly involved in NTH α1(IV) production. NTH electrophoretic mobility on SDS-PAGE was higher under hypoxia or deferoxamine treatment, likely due to suppressed lysyl hydroxylase 3 activity. This study demonstrates NTH production in tumor cells and tissues under hypoxia, suggesting their association with tumor angiogenesis and potential as therapeutic targets.","PeriodicalId":15234,"journal":{"name":"Journal of biochemistry","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2025-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142931804","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}

引用次数: 0

Polymerase-Usage Sequencing Identifies Initiation Zones with Less Bias Across S phase in Mouse Embryonic Stem Cells. 聚合酶使用测序鉴定小鼠胚胎干细胞S期起始区偏差较小。

IF 2.1 4区生物学

Journal of biochemistry Pub Date : 2025-01-02 DOI: 10.1093/jb/mvae097

Akino Matsumoto, Yasukazu Daigaku, Tomomi Tsubouchi

引用次数: 0