• Title/Summary/Keyword: Protein Function

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Protein Aggregation and Adsorption upon In vitro Refolding of Recombinant Pseudomonas Lipase

  • Lee, Young-Phil;Rhee, Joon-Shick
    • Journal of Microbiology and Biotechnology
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    • v.6 no.6
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    • pp.456-460
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    • 1996
  • Recombinant Pseudomonas lipase was used to study protein aggregation and adsorption upon in vitro refolding. Protein adsorption as well as aggregation was responsible for major side reactions upon in vitro refolding as a function of protein concentration. The optimal range of protein concentration was determined by the relative contribution of protein aggregation and adsorption. Above the optimal range, the yield of active lipase inversely correlated with protein aggregation, showing a competition between folding and aggregation. However, adsorption of protein rather than protein aggregation is thought to contribute as a major side reaction of the refolding process at sub-optimal concentrations at which the formation of aggregates should be more reduced. Protein aggregation was influenced by the amount of guanidine hydrochloride in the refolding solvent. The refolding temperature was a critical factor determining the extent of protein aggregation. The refolding yield was also affected by the dilution fold and dilution mode, which suggests that the refolding process might kinetically compete with the rate of mixing.

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Correlation between Body Composition and Lung Function in Healthy Adults (정상 성인의 신체조성과 폐 기능의 연관성)

  • Kim, Hyunseung;Cho, Sunghyoun
    • Journal of The Korean Society of Integrative Medicine
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    • v.8 no.2
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    • pp.53-61
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    • 2020
  • Purpose : We investigated the correlation between body composition and lung function in healthy adults. Methods : This study included 204 healthy adults in whom all measurements were obtained once, and all data were analyzed using the SPSS software for Windows, version 22.0. Pearson's correlation analysis was performed to determine the correlation between body composition (represented by the total body water, protein mass, soft lean mass, mineral mass, basal metabolic rate, fat-free mass, skeletal muscle mass, and body fat percentage) and lung function (represented by the forced vital capacity [FVC], forced expiratory volume in 1 second [FEV1], the FEV1/FVC ratio, maximum voluntary ventilation [MVV], maximum expiratory pressure [MEP], and the maximum inspiratory pressure [MIP]). All measurements were obtained by two investigators to improve reliability. A significance level of α=.05 was used to verify statistical significance. Results : Among the lung function measurements obtained in both men and women, the FVC, FEV1, MVV, and MIP were positively correlated with the total body water, protein mass, soft lean mass, mineral mass, basal metabolic rate, fat-free mass, and skeletal muscle mass in men (p<.05). The FEV1/FVC ratio was negatively correlated with the total body water, soft lean mass, mineral mass, basal metabolic rate, fat-free mass and the body fat percentage (p<.05). Notably, the FVC, FEV1, and MVV were positively correlated with the total body water, protein mass, soft lean mass, mineral mass, basal metabolic rate, fat-free mass, and skeletal muscle mass in women (p<.05). Conclusion : This study showed a significant correlation between body composition and lung function in healthy adults. In combination with future studies on lung function, our results can provide objective evidence regarding the importance of prevention of lung disease, and our data can be utilized in rehabilitation programs for patients with respiratory diseases.

Analysis of a Large-scale Protein Structural Interactome: Ageing Protein structures and the most important protein domain

  • Bolser, Dan;Dafas, Panos;Harrington, Richard;Schroeder, Michael;Park, Jong
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2003.10a
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    • pp.26-51
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    • 2003
  • Large scale protein interaction maps provide a new, global perspective with which to analyse protein function. PSIMAP, the Protein Structural Interactome Map, is a database of all the structurally observed interactions between superfamilies of protein domains with known three-dimensional structure in thePDB. PSIMAP incorporates both functional and evolutionary information into a single network. It makes it possible to age protein domains in terms of taxonomic diversity, interaction and function. One consequence of it is to predict the most important protein domain structure in evolution. We present a global analysis of PSIMAP using several distinct network measures relating to centrality, interactivity, fault-tolerance, and taxonomic diversity. We found the following results: ${\bullet}$ Centrality: we show that the center and barycenter of PSIMAP do not coincide, and that the superfamilies forming the barycenter relate to very general functions, while those constituting the center relate to enzymatic activity. ${\bullet}$ Interactivity: we identify the P-loop and immunoglobulin superfamilies as the most highly interactive. We successfully use connectivity and cluster index, which characterise the connectivity of a superfamily's neighbourhood, to discover superfamilies of complex I and II. This is particularly significant as the structure of complex I is not yet solved. ${\bullet}$ Taxonomic diversity: we found that highly interactive superfamilies are in general taxonomically very diverse and are thus amongst the oldest. This led to the prediction of the oldest and most important protein domain in evolution of lift. ${\bullet}$ Fault-tolerance: we found that the network is very robust as for the majority of superfamilies removal from the network will not break up the network. Overall, we can single out the P-loop containing nucleotide triphosphate hydrolases superfamily as it is the most highly connected and has the highest taxonomic diversity. In addition, this superfamily has the highest interaction rank, is the barycenter of the network (it has the shortest average path to every other superfamily in the network), and is an articulation vertex, whose removal will disconnect the network. More generally, we conclude that the graph-theoretic and taxonomic analysis of PSIMAP is an important step towards the understanding of protein function and could be an important tool for tracing the evolution of life at the molecular level.

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Expression of Heat Shock Protein HspA2 in Human Tissues (인간 조직에서 Heat Shock Protein A2 (HspA2) 단백질의 발현)

  • Son, W.Y.;Hwang, S.H.;Han, C.T.;Lee, J.H.;Choi, Y.J.;Kim, S.;Kim, Y.C.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.2
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    • pp.225-230
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    • 1999
  • In mouse, the heat shock protein 70-2 (hsp70-2) is found to have special function in spermatogenesis. Based on the observation, the hypothesis that human hspA2 (human gene; 98.2% amino acid homology with hsp70-2) might have important function in spermatogenesis in human testes was proposed. To test the hypothesis, we examined the expression of hspA2 in human tissues. Expression vector pDMC4 for expression of the human hspA2 protein using pTricHisB (invitrogen, USA) was constructed and the expressed hspA2 protein was cross-reacted with antiserum 2A raised against mouse hsp70-2 protein. Based on the cross-reactivity, we determined the expression level of hspA2 protein in human tissues by western blot analysis using the antiserum 2A. We demonstrated that antiserum 2A antibodies detected human hspA2 protein with specificity which was produced in the E.coli expression system. On Western blot analyses, significant hspA2 expression was observed in testes with normal spermatogenesis, whereas a low level of hspA2 was expressed in testis with Sertoli-cell only syndrome. Also, a small amount of hspA2 was detected in breast, stomach, prostate, colon, liver, ovary, and epididymis. These results demonstrate that the hspA2 protein is highly expressed in male specific germ cells, which in turn suggests that hspA2 protein might playa specific role during meiosis in human testes as suggested in the murine model. However, further studies should be attempted to determine the function of hspA2 protein in human spermatogenesis.

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Acetohydroxyacid Synthase

  • Duggleby, Ronald G.;Pang, Siew Siew
    • BMB Reports
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    • v.33 no.1
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    • pp.1-36
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    • 2000
  • Acetohydroxyacid synthase (EC 4.1.3.18) catalyses the first reaction in the pathway for synthesis of the branched-chain amino acids. The enzyme is inhibited by several commercial herbicides and has been subjected to detailed study over the last 20 to 30 years. Here we review the progress that has been made in understanding its structure, regulation, mechanism, and inhibition.

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Proline Metabolism in Neurological and Psychiatric Disorders

  • Yao, Yuxiao;Han, Weiping
    • Molecules and Cells
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    • v.45 no.11
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    • pp.781-788
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    • 2022
  • Proline plays a multifaceted role in protein synthesis, redox balance, cell fate regulation, brain development, and other cellular and physiological processes. Here, we focus our review on proline metabolism in neurons, highlighting the role of dysregulated proline metabolism in neuronal dysfunction and consequently neurological and psychiatric disorders. We will discuss the association between genetic and protein function of enzymes in the proline pathway and the development of neurological and psychiatric disorders. We will conclude with a potential mechanism of proline metabolism in neuronal function and mental health.

Effect of blended protein nutritional support on reducing burn-induced inflammation and organ injury

  • Yu, Yonghui;Zhang, Jingjie;Wang, Jing;Wang, Jing;Chai, Jiake
    • Nutrition Research and Practice
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    • v.16 no.5
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    • pp.589-603
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    • 2022
  • BACKGROUND/OBJECTIVES: Previous studies have reported that protein supplementation contributes to the attenuation of inflammation. Serious trauma such as burn injury usually results in the excessive release of inflammatory factors and organs dysfunction. However, a few reports continued to focus on the function of protein ingestion in regulating burn-induced inflammation and organ dysfunction. MATERIALS/METHODS: This study established the rat model of 30% total body surface area burn injury, and evaluated the function of blended protein (mixture of whey and soybean proteins). Blood routine examination, inflammatory factors, blood biochemistry, and immunohistochemical assays were employed to analyze the samples from different treatment groups. RESULTS: Our results indicated a decrease in the numbers of white blood cells, monocytes, and neutrophils in the burn injury group administered with the blended protein nutritional support (Burn+BP), as compared to the burn injury group administered normal saline supplementation (Burn+S). Expressions of the pro-inflammatory factors (tumor necrosis factor-α and interleukin-6 [IL-6]) and chemokines (macrophage chemoattractant protein-1, regulated upon activation normal T cell expressed and secreted factor, and C-C motif chemokine 11) were dramatically decreased, whereas anti-inflammatory factors (IL-4, IL-10, and IL-13) were significantly increased in the Burn+BP group. Kidney function related markers blood urea nitrogen and serum creatinine, and the liver function related markers alanine transaminase, aspartate aminotransferase, alkaline phosphatase, and lactate dehydrogenase were remarkably reduced, whereas albumin levels were elevated in the Burn+BP group as compared to levels obtained in the Burn+S group. Furthermore, inflammatory cells infiltration of the kidney and liver was also attenuated after burn injury administered with blended protein supplementation. CONCLUSIONS: In summary, nutritional support with blended proteins dramatically attenuates the burn-induced inflammatory reaction and protects organ functions. We believe this is a new insight into a potential therapeutic strategy for nutritional support of burn patients.

Effect of dietary protein level on bone metabolism of young and aged rats (식이 단백질 수준이 어린쥐와 나이든 쥐의 골격의 대사에 미치는 영향)

  • 조미숙
    • Journal of Nutrition and Health
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    • v.22 no.6
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    • pp.497-506
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    • 1989
  • To investigate the effect of levels of dietary protein and age on bone metabolism 40% and 5% casein were fed to the rats of 2 & 13 months of age for 12 weeks. High protein groups showed higher bone weight and Ca content than low protein groups and urinary Ca loss was increased in high protein groups but the difference disappeared gradually. A significant increase in urinary hydroxyproline excretion was noted in high protein groups of both age. Another short term study was undertaken to study if the above effect was related with renal function or PTH. Extremely high and low protein diets(60%, 6%) were fed to the rats of different ages(6wks, 6mos.) for 2 weeks, Urinary Ca excretion was significantly increased in high protein groups of young and aged rats and GFR was increased as well. There was no difference in serum iPTH levels between low and high protein groups, but it was elevated in aged rats. Alkaline phosphatase activity was higher in young rats, reflecting faster bone formation. The observed hypercalciuria in high protein groups, especially in aged rats, seems to be related to higher GFR, and PTH dose not appear to be a major mediator.

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Expression of PDL-specific protein;PDLs22 on the developing mouse tooth and periodontium (발생중인 생쥐 치아 및 치주조직에서 치주인대-특이 단백질; PDLs22의 발현)

  • Park, Jung-Won;Park, Byung-Ki;Kim, Sang-Mok;Kim, Byung-Ock;Park, Joo-Cheol
    • Journal of Periodontal and Implant Science
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    • v.32 no.1
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    • pp.1-12
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    • 2002
  • The periodontal ligament(PDL) is a unique tissue that is crucial for tooth function. However, little is known of the molecular mechanisms controlling PDL function. PDL-specific protein;PDLs22 had been previously identified as a novel protein isolated from cultured human PDL fibroblasts using subtraction hybridization between human gingival fibroblasts and PDL fibroblasts. The aim of this study was to examine the expression pattern and tissue localization of PDLs22 protein in embryonic and various postnatal stages of developing mouse using immunohistochemical staining. Embryos (E18) and postnatal (P1, P4, P5, P15, P18) were decapitated and the heads were fixed overnight in a freshly prepared solution of 4% paraformaldehyde. Some specimens were decalcified for $2{\sim}4$ weeks in a solution containing 10% of the disodium salt of ethylenediamine-tetraacetic acid (EDTA). Next, tissues were dehydrated, embedded in paraffin and sectioned serially at $6{\mu}m$ in thickness. Polyclonal antiserum raised against PDLs22 peptides, ISNKYLVKRQSRD, were made. The localization of PDLs22 in tissues was detected by polyclonal antibody against PDLs22 by means of immunohistochemical staining. The results were as follows; 1. Expression of PDLs22 protein was not detected in the tooth germ of bud and cap stage. 2. At the late bell stage and root formation stage, strong expression of PDLs22 protein was observed in developing tooth follicle, osteoblast-like cells, and subodontoblastic cells in the tooth pulp, but not in gingival fibroblasts, ameloblasts and odontoblasts of tooth germ 3. In erupted tooth, PDLs22 protein was intensely expressed in PDL and osteoblast-like cells of alveolar bone, but not in gingival fibroblasts, mature osteocytes and adjacent salivary glands. 4. In the developing alveolar bone and mid-palatal suture, expression of PDLs22 protein was seen in undifferentiated mesenchymal cells and osteoblast-like cells of developing mid-palatal suture, but not in mature osteocytes and chondrocytes. These results suggest that PDLs22 protein may play an important role in the differentiation of undifferentiated mesenchymal cells in the bone marrow and PDL cells, which can differentiate into multiple cell types including osteoblasts, cementoblasts, and PDL fibroblasts. However, more researches should be performed to gain a better understanding of the exact function of PDLs22 protein which related to the PDL cell differentiation.