The formation of most proteins consists of two steps: the synthesis of precursor proteins and the synthesis of functional proteins. In these processes, propeptides play important roles in assisting protein folding or inhibiting its activity. As an important polypeptide chain coded by a gene sequence in lipase gene, propeptide usually functions as an intramolecular chaperone, assisting enzyme molecule folding. Meanwhile, some specific sites on propeptide such as glycosylated sites, have important effect on the activity, stability in extreme environment, methanol resistance and the substrate specificity of the lipase. Studying the mechanism of propeptide-mediated protein folding, as well as the influence of propeptide on lipases, will allow to regulate lipase by alternating the propeptide folding behavior and in turn pave new ways for protein engineering research.
Lipase/metabolism* ; Molecular Chaperones/metabolism* ; Protein Folding ; Protein Precursors ; Substrate Specificity

protein spots to be differentially expressed by more than 2-fold change with p<0.05 considered as significant. Matrix-assisted laser desorption/ionization time of flight/mass spectrometry identified GPx4, JIP4, ZN248 to be overexpressed while HSPA2, GSTM5, TF3C1, CC74A was underexpressed in RPL group. Western blot analysis confirmed the differential expression of key redox associated proteins GPx4 and HSPA2 in the RPL group. Functional analysis revealed the involvement of key biological processes that includes spermatogenesis, response to oxidative stress, protein folding and metabolic process.CONCLUSIONS: The present study provides a snapshot of the altered protein expression levels consistent with the potential involvement of the sperm chromatin landscape in early embryonic development.]]>
Abortion, Spontaneous ; Biological Processes ; Blotting, Western ; Chromatin ; Embryo Loss ; Embryonic Development ; Female ; Healthy Volunteers ; Humans ; Karyotyping ; Male ; Masturbation ; Metabolism ; Oxidation-Reduction ; Oxidative Stress ; Pregnancy ; Prospective Studies ; Protein Folding ; Proteomics ; Sexual Abstinence ; Spectrum Analysis ; Spermatogenesis ; Spermatozoa ; Two-Dimensional Difference Gel Electrophoresis ; World Health Organization

Artificial intelligence (AI), big data, and ubiquitous robotic companions —the three most notable technologies of the 4th Industrial Revolution—are receiving renewed attention each day. Technologies that can be experienced in daily life, such as autonomous navigation, real-time translators, and voice recognition services, are already being commercialized in the field of information technology. In the biosciences field in Korea, such technologies have become known to the local public with the introduction of the AI doctor Watson in large number of hospitals. Additionally, AlphaFold, a technology resembling the AI AlphaGo for the game Go, has surpassed the limit on protein folding predictions—the most challenging problems in the field of protein biology. This report discusses the significance of AI technology and big data on the bioscience field. The introduction of automated robots in this field is not just only for the purpose of convenience but a prerequisite for the real sense of AI and the consequent accumulation of basic scientific knowledge.
Artificial Intelligence ; Biology ; Biotechnology ; Friends ; Humans ; Korea ; Protein Folding ; Voice

Rotavirus (RV)-infected piglets are presumed to be latent sources of heterologous RV infection in humans and other animals. In RVs, non-structural protein 4 (NSP4) is the major virulence factor with pleiotropic properties. In this study, we analyzed the nsp4 gene from porcine RVs isolated from diarrheic and non-diarrheic cases at different levels of protein folding to explore correlations to diarrhea-inducing capabilities and evolution of nsp4 in the porcine population. Full-length nsp4 genes were amplified, cloned, sequenced, and then analyzed for antigenic epitopes, RotaC classification, homology, genetic relationship, modeling of NSP4 protein, and prediction of post-translational modification. RV presence was observed in both diarrheic and non-diarrheic piglets. All nsp4 genes possessed the E1 genotype. Comparison of primary, secondary, and tertiary structure and the prediction of post-translational modifications of NSP4 from diarrheic and non-diarrheic piglets revealed no apparent differences. Sequence analysis indicated that nsp4 genes have a multi-phyletic evolutionary origin and exhibit species independent genetic diversity. The results emphasize the evolution of the E9 nsp4 genotype from the E1 genotype and suggest that the diarrhea-inducing capability of porcine RVs may not be exclusively linked to its enterotoxin gene.
Animals ; Classification ; Clone Cells ; Enterotoxins ; Epitopes ; Genetic Variation ; Genotype ; Humans ; Protein Folding ; Protein Processing, Post-Translational ; Rotavirus ; Sequence Analysis ; Viral Nonstructural Proteins ; Virulence

Free fatty acids (FFAs) are important substrates for mitochondrial oxidative metabolism and ATP synthesis but also cause serious stress to various tissues, contributing to the development of metabolic diseases. CD36 is a major mediator of cellular FFA uptake. Inside the cell, saturated FFAs are able to induce the production of cytosolic and mitochondrial reactive oxygen species (ROS), which can be prevented by co-exposure to unsaturated FFAs. There are close connections between oxidative stress and organellar Ca²⁺ homeostasis. Highly oxidative conditions induced by palmitate trigger aberrant endoplasmic reticulum (ER) Ca²⁺ release and thereby deplete ER Ca²⁺ stores. The resulting ER Ca²⁺ deficiency impairs chaperones of the protein folding machinery, leading to the accumulation of misfolded proteins. This ER stress may further aggravate oxidative stress by augmenting ER ROS production. Secondary to ER Ca²⁺ release, cytosolic and mitochondrial matrix Ca²⁺ concentrations can also be altered. In addition, plasmalemmal ion channels operated by ER Ca²⁺ depletion mediate persistent Ca²⁺ influx, further impairing cytosolic and mitochondrial Ca²⁺ homeostasis. Mitochondrial Ca²⁺ overload causes superoxide production and functional impairment, culminating in apoptosis. This vicious cycle of lipotoxicity occurs in multiple tissues, resulting in β-cell failure and insulin resistance in target tissues, and further aggravates diabetic complications.
Adenosine Triphosphate ; Apoptosis ; Calcium* ; Cytosol ; Diabetes Complications ; Endoplasmic Reticulum ; Fatty Acids, Nonesterified ; Homeostasis ; Insulin Resistance ; Ion Channels ; Metabolic Diseases ; Metabolism ; Oxidative Stress* ; Protein Folding ; Reactive Oxygen Species ; Superoxides

The endoplasmic reticulum (ER) is an important subcellular organelle that is involved in numerous activities required to achieve and maintain functional proteins in addition to its role in the biosynthesis of lipids and as a repository of intracellular Ca²⁺. The inability of the ER to cope with protein folding beyond its capacity causes disturbances that evoke ER stress. Cells possess molecular mechanisms aimed at clearing unwanted cargo from the ER lumen as an adaptive response, but failing to do so navigates the system towards cell death. This systemic approach is called the unfolded protein response. Aging insults cells through various perturbations in homeostasis that involve curtailing ER function by mitigating the expression of its resident chaperones and enzymes. Here the unfolded protein response (UPR) cannot protect the cell due to the weakening of its protective arm, which exacerbates imbalanced homeostasis. Aging predisposed breast malignancy activates the UPR, but tumor cells maneuver the mechanistic details of the UPR, favoring tumorigenesis and thereby eliciting a treacherous condition. Tumor cells exploit UPR pathways via crosstalk involving various signaling cascades that usher tumor cells to immortality. This review aims to present a collection of data that can delineate the missing links of molecular signatures between aging and breast cancer.
Aging* ; Arm ; Breast Neoplasms* ; Breast* ; Carcinogenesis ; Cell Death ; Endoplasmic Reticulum* ; Homeostasis ; Organelles ; Protein Folding ; Unfolded Protein Response*

Excessive production of reactive oxygen species (ROS) and endoplasmic reticulum (ER) stress-mediated responses are critical to embryonic development in the challenging in vitro environment. ROS production increases during early embryonic development with the increase in protein requirements for cell survival and growth. The ER is a multifunctional cellular organelle responsible for protein folding, modification, and cellular homeostasis. ER stress is activated by a variety of factors including ROS. Such stress leads to activation of the adaptive unfolded protein response (UPR), which restores homeostasis. However, chronic stress can exceed the toleration level of the ER, resulting in cellular apoptosis. In this review, we briefly describe the generation and impact of ROS in preimplantation embryo development, the ROS-mediated activation mechanism of the UPR via the ER, and the subsequent activation of signaling pathways following ER stress in preimplantation embryos.
Apoptosis ; Blastocyst* ; Cell Survival ; Embryonic Development ; Endoplasmic Reticulum ; Female ; Homeostasis ; In Vitro Techniques ; Organelles ; Oxygen* ; Pregnancy ; Protein Folding ; Reactive Oxygen Species ; Unfolded Protein Response*

Effective expression of pIFN-α in recombinant Pichia pastoris was conducted in a 5 L fermentor. Ethanol accumulation during the late glycerol feeding period inhibited heterologous protein expression. Comparative transcriptome analysis was thus performed to compare the gene transcription profiles of Pichia pastoris KM71H in high and low ethanol concentration environments. The results showed that during the glycerol cultivation stage, 545 genes (265 up-regulated and 280 down-regulated) were differentially expressed with ethanol stress. These genes were mainly involved in protein synthesis, energy metabolism, cell cycle and peroxisome metabolism. During the methanol induction stage, 294 genes (171 up-regulated and 123 down-regulated) were differentially expressed, which were mainly related to methanol metabolism, amino acid metabolism and protein synthesis. Ethanol stress increased protein misfolding and reduced structural integrity of ribosome and mitochondria during cultivation stage, and led to the failure of endoplasmic reticulum stress removal and damaged amino acid metabolism during induction stage in Pichia pastoris.
Amino Acids ; metabolism ; Bioreactors ; Endoplasmic Reticulum Stress ; Energy Metabolism ; Ethanol ; chemistry ; Gene Expression Profiling ; Gene Expression Regulation, Fungal ; Glycerol ; Methanol ; Pichia ; metabolism ; Protein Biosynthesis ; drug effects ; Protein Folding ; Recombinant Proteins ; biosynthesis ; Transcriptome

Influenza C virus, a member of the Orthomyxoviridae family, causes flu-like disease but typically only with mild symptoms. Humans are the main reservoir of the virus, but it also infects pigs and dogs. Very recently, influenza C-like viruses were isolated from pigs and cattle that differ from classical influenza C virus and might constitute a new influenza virus genus. Influenza C virus is unique since it contains only one spike protein, the hemagglutinin-esterase-fusion glycoprotein HEF that possesses receptor binding, receptor destroying and membrane fusion activities, thus combining the functions of Hemagglutinin (HA) and Neuraminidase (NA) of influenza A and B viruses. Here we briefly review the epidemiology and pathology of the virus and the morphology of virus particles and their genome. The main focus is on the structure of the HEF protein as well as on its co- and post-translational modification, such as N-glycosylation, disulfide bond formation, S-acylation and proteolytic cleavage into HEF1 and HEF2 subunits. Finally, we describe the functions of HEF: receptor binding, esterase activity and membrane fusion.
Animals ; Cattle ; Dogs ; Hemagglutinins, Viral ; chemistry ; metabolism ; Influenzavirus C ; physiology ; Orthomyxoviridae Infections ; metabolism ; virology ; Protein Conformation ; Protein Folding ; Protein Processing, Post-Translational ; Viral Fusion Proteins ; chemistry ; metabolism

OBJECTIVETo study the drug sensitivity and analyze the replication kinetics of HIV-1 B and CRF07_BC subtypes with I132L or T139K/R mutations.

METHODSThe amino acids in position 132 and 139 of reverse transcriptase (RT) region of the infectious clone PNL4-3 (HIV-1 B subtype) were changed to L and T/R through site mutagenesis. Combined with the previously constructed infectious clone of HIV-1 CRF07_BC subtype with I132L and T139K/R mutations in RT region, mutated PNL4-3 infectious clones were transfected into 293T cells. The infection ability of mutated clones was detected. The drug sensitivity to NNRTIs (TMC-125, DLV, NVP, EFV) and the properties of replication kinetics were also evaluated.

RESULTSThe mutated infectious clones were constructed including PNL4-3-RT-I132L, PNL4-3-RT-T139K and PNL4-3-RT-T139R. The I132L and T139K/R mutations in HIV-1 B and CRF07_BC infectious clones reduced their drug sensitivity to NNRTIs, which accompanied with the increase of EC50 (concentration for 50% of maximal effect). In subtype CRF07_BC, I132L mutation increased EC50 by 2.55, 19.35, 28.05, 6.13 fold, T139K mutation increased EC50 by 4.67, 3.66, 7.35, 3.30 fold, and T139R mutation increased EC50 by 1.82, 4.69, 25.12, 1.89 fold, respectively. In subtype B, I132L increased EC50 by 3.91, 4.61, 6.38, 3.56 fold, T139K increased EC50 by 3.13, 1.78, 2.26, 2.10 fold, T139R increased EC50 by 5.79, 3.99, 5.78, 2.75 fold, respectively. Similar as wild type PNL4-3, the replication ability of 132L/139K/139R mutated infectious clones reached the peak in day 11. However, compared to wild type BC-WT, I132L/T139R mutations delayed the peak time to day 14 and 21.

CONCLUSIONThe novel drug resistance associated mutations I132L and T139K/R can reduce the drug sensitivity to NNRTIs in subtype B and CRF07_BC, and the replication ability of CRF_07BC declined by I132L mutation.

Anti-HIV Agents ; Drug Resistance ; Genotype ; HIV-1 ; Kinetics ; Mutation ; Polymorphism, Single Nucleotide ; Protein Folding ; Pyridazines ; Reverse Transcriptase Inhibitors ; Virus Replication