Effects of osmolytes on Pelodiscus sinensis creatine kinase: A study on thermal denaturation and aggregation

Wang, Wei; Lee, Jinhyuk; Jin, Qin-Xin; Fang, Nai-Yun; Si, Yue-Xiu; Yin, Sheng; Qian, Guo‐Ying; Park, Yong‐Doo

doi:10.1016/j.ijbiomac.2013.06.015

Cited by 17 publications

(6 citation statements)

References 30 publications

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“…TMAO is considered as one of the strongest stabilizers in nature, and participate in the protection of renal cells of mammals from deleterious influence of urea (Jackson-Atogi et al, 2013;Sarma & Paul, 2013), or stabilization of the cells of deep see organisms against high pressure (Sarma & Paul, 2013;Sarma & Paul, 2012). Glycine, NMG, DMG, and TMG (Betaine) are generally considered as moderate stabilizers (Santoro et al, 1992;Wang et al, 2013;Guinn et al, 2011;Caldas et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

Protein thermal stabilization in aqueous solutions of osmolytes.

et al. 2015

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Proteins' thermal stabilization is a significant problem in various biomedical, biotechnological, and technological applications. We investigated thermal stability of hen egg white lysozyme in aqueous solutions of the following stabilizing osmolytes: Glycine (GLY), N-methylglycine (NMG), N,N-dimethylglycine (DMG), N,N,N-trimethylglycine (TMG), and trimethyl-N-oxide (TMAO). Results of CD-UV spectroscopic investigation were compared with FTIR hydration studies' results. Selected osmolytes increased lysozyme's thermal stability in the following order: Gly>NMG>TMAO≈DMG>TMG. Theoretical calculations (DFT) showed clearly that osmolytes' amino group protons and water molecules interacting with them played a distinctive role in protein thermal stabilization. The results brought us a step closer to the exact mechanism of protein stabilization by osmolytes.

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Section: Introductionmentioning

confidence: 99%

Protein thermal stabilization in aqueous solutions of osmolytes.

et al. 2015

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“…), most of the enzyme would be aggregated by heating at 50°C (Wang et al . ) and the amount of the enzyme solubilized into 0.2 mol/L NaCl seems to be small. Thus, even if a weak band around 44.3 kDa of sample heated for 0 min (lane f in Fig.…”

Section: Resultsmentioning

confidence: 89%

Characterization of myofibrillar adenosine triphosphatase activity and liberation of actin from myofibrils upon heating chicken breast meat

Matsuishi

Eda

Saito

et al. 2017

Animal Science Journal

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Denaturation of actin and myosin in myofibrils induced by heating at 50°C was investigated to reveal the mechanism of irreversible liberation of actin from myofibrils on heating at lower temperatures than conventional cooking. Denaturation of these proteins was determined by Mg -ATPase (adenosine triphosphatase) and Ca -ATPase activities. When minced meat was heated for 20 min, actin was liberated accompanying denaturation of 80% of actin and 50% of myosin. Heating of the myofibrillar fraction (MFF) isolated from meat homogenate induced much slower denaturation of actin than myosin. When MFF was heated with sarcoplasmic fractions, denaturation of actin was facilitated, suggesting that sarcoplasmic fractions contain factors to facilitate actin denaturation. Inosine-5'-monophosphate, a component of sarcoplasmic fractions, was shown to have no effect on actin and myosin denaturation. These results suggest that heating meat at 50°C dissociates binding ('Bond A') between actin and myosin participating in ATPase activities, resulting in denaturation of both proteins under influence of sarcoplasmic components. Although denaturation of actin and myosin disrupted Bond A, actin was not liberated simultaneously, suggesting the presence of another bond ('Bond B', more heat-stable than Bond A) between both proteins and necessity of disruption of Bond B for actin release from myofibrils.

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“…The crystallographic protein structures for PSCK isozymes have not yet been elucidated. Therefore we constructed the 3D structures by homology modeling according to previous reports [15,16,20,21]. Each PSCK-B, PSCK-M, and PSCK-S protein structure was modeled using homology templates HHsearch [22] to find template proteins, and the 9 or 10 best template sequences were aligned by application of T-Coffee [23] in its fast mode.…”

Section: Computational Docking Simulations and Molecular Dynamics (Md)mentioning

confidence: 99%

“…The supernatant was obtained by freezing centrifugation at 5000 rpm for 15 min for biochemical parameters' assay. The total activity of PSCK from the tissues was measured following proton generation during the reaction of ATP and creatine with thymol blue at 597 nm and 25°C, as previously described [15,16,29,30]. The activities of total superoxide dismutase (T-SOD), lactate dehydrogenase assay (LDH), malondialdehyde (MDA), catalase (CAT), and the content of ATP were measured according to the kit manuals (A020-2 for LDH, A001-1 for T-SOD, A007-1 for CAT, A003-2 for MDA, and A095 for ATP) provided by Nanjing Jiancheng Biochemical Reagent Co. Ltd. (Nanjing, China) using the clinical biochemical indicator autoanalyzer: Infinite M200Pro spectrometer (Tecan, Switzerland), according to the manufacturer's instructions.…”

Section: Determination Of Biochemical Enzyme Activitiesmentioning

confidence: 99%

“…Previously we have reported two isoforms of CK from P. sinensis (PSCK-B and PSCK-M) -GenBank accession No. JQ410386.1 and KR633145.1 respectively [15,16], analyzed by RACE technique. As a sequential study, we have analyzed the mitochondrial isoform of P. sinensis identified as type II mitochondrial isoform (PSCK-S), listed in Supplementary Table 1.…”

Section: Introductionmentioning

confidence: 99%

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Comparative studies of the expression of creatine kinase isoforms under immune stress in Pelodiscus sinensis

Wang

Lee

et al. 2020

International Journal of Biological Macromolecules

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The expression and localization of different isoforms of creatine kinase in Pelodiscus sinensis (PSCK) were studied to reveal the role of PSCK isozymes (PSCK-B, PSCK-M, PSCK-S) under bacterial infection-induced immunologic stress. The computational molecular dynamics simulations predicted that PSCK-S would mostly possess a kinase function in a structural aspect when compared to PSCK-B and PSCK-M. The assay of biochemical parameters such as total superoxide dismutase (T-SOD), lactate dehydrogenase (LDH), malondialdehyde (MDA), catalase (CAT), and the content of ATP were measured along with total PSCK activity in different tissue samples under bacterial infection. The expression detections of PSCK isozymes in vitro and in vivo were overall well-matched where PSCK isozymes were expressed differently in P. sinensis tissues. The results showed that PSCK-B mostly contributes to the spleen, followed by the liver and myocardium; PSCK-M mostly contributes to the liver, followed by the myocardium and skeletal muscle, while PSCK-S contributes to the spleen and is uniquely expressed in skeletal muscle. Our study suggests that the various alterations of PSCK isozymes in tissues of P. sinensis are prone to defense the bacterial infection and blocking energetic imbalance before severe pathogenesis turned on in P. sinensis.

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Effects of osmolytes on Pelodiscus sinensis creatine kinase: A study on thermal denaturation and aggregation

Cited by 17 publications

References 30 publications

Protein thermal stabilization in aqueous solutions of osmolytes.

Protein thermal stabilization in aqueous solutions of osmolytes.

Characterization of myofibrillar adenosine triphosphatase activity and liberation of actin from myofibrils upon heating chicken breast meat

Comparative studies of the expression of creatine kinase isoforms under immune stress in Pelodiscus sinensis

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