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Point Mutations in the Split PLC-γ1 PH Domain Modulate Phosphoinositide Binding
( Sung Kuk Kim ),( Sung Mo Wee ),( Jong Soo Chang ),( Taeg Kyu Kwon ),( Do Sik Min ),( Young Han Lee ),( Pann Ghill Suh ) 생화학분자생물학회 2004 BMB Reports Vol.37 No.6
A number of signaling molecules contain small pleckstrin homology (PH) domains capable of binding phosphoinositides or proteins. Phospholipase C (PLC)-γ1 has two putative PH domains, an NH₂-terminal (PH1) and a split PH domain (nPH₂ and cPH₂). We previously reported that the split PH domain of PLC-γ1 binds to phosphatidγ1inositol 4-phosphate (PI(4)P) and phosphatidγ1inositol 4,5-bisphosphate (PI(4,5)P₂) (Chang et al., 2002). To identify the amino acid residues responsible for binding with PI(4)P and PI(4,5)P₂, we used site-directed mutagenesis to replace each amino acid in the variable loop-1 (VL-l) region of the PLC-γ1 nPH₂ domain with alanine (a neutral amino acid). The phosphoinositide-binding affinity of these mutant molecules was analyzed by Dot-blot assay followed by ECL detection. We found that two PLC-γ1 nPH2 domain mutants, P500A and HSO3A, showed reduced affinities for phosphoinositide binding. Furthermore, these mutant PLC-γ1 molecules showed reduced PI(4,5)P₂ hydrolysis. Using green fluorescent protein (GFP) fusion protein system, we showed that both PH₁ and nPH₂ domains are responsible for membrane-targeted translocation of PLC-γ1 upon serum stimulation. Together, our data reveal that the amino acid residues Pro^(500) and His^(503) are critical for binding of PLC-γ1 to one of its substrates, PI(4,5 )P₂ in the membrane.
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Mo, SangJoon,Lee, Sung-Kwon,Jin, Ying-Yu,Oh, Chung-Hun,Suh, Joo-Won Springer-Verlag 2013 Applied microbiology and biotechnology Vol.97 No.7
<P>FK506 production by a mutant strain (Streptomyces sp. RM7011) induced by N-methyl-N'-nitro-N-nitrosoguanidine and ultraviolet mutagenesis was improved by 11.63-fold (94.24?mg/l) compared to that of the wild-type strain. Among three different metabolic pathways involved in the biosynthesis of methylmalonyl-CoA, only expression of propionyl-CoA carboxylase (PCC) pathway led to a 1.75-fold and 2.5-fold increase in FK506 production and the methylmalonyl-CoA pool, respectively, compared to those of the RM7011 strain. Lipase activity of the high FK506 producer mutant increased in direct proportion to the increase in FK506 yield, from low detection level up to 43.1?U/ml (12.6-fold). The level of specific FK506 production and lipase activity was improved by enhancing the supply of lipase inducers. This improvement was approximately 1.88-fold (71.5?mg/g) with the supplementation of 5?mM Tween 80, which is the probable effective stimulator in lipase production, to the R2YE medium. When 5?mM vinyl propionate was added as a precursor for PCC pathway to R2YE medium, the specific production of FK506 increased approximately 1.9-fold (71.61?mg/g) compared to that under the non-supplemented condition. Moreover, in the presence of 5?mM Tween 80, the specific FK506 production was approximately 2.2-fold (157.44?mg/g) higher than that when only vinyl propionate was added to the R2YE medium. In particular, PCC expression in Streptomyces sp. RM7011 (RM7011/pSJ1003) together with vinyl propionate feeding resulted in an increase in the FK506 titer to as much as 1.6-fold (251.9?mg/g) compared with that in RM7011/pSE34 in R2YE medium with 5?mM Tween 80 supplementation, indicating that the vinyl propionate is more catabolized to propionate by stimulated lipase activity on Tween 80, that propionyl-CoA yielded from propionate generates methylmalonyl-CoA, and that the PCC pathway plays a key role in increasing the methylmalonyl-CoA pool for FK506 biosynthesis in RM7011 strain. Overall, these results show that a combined approach involving classical random mutation and metabolic engineering can be applied to supply the limiting factor for FK506 biosynthesis, and vinyl propionate could be successfully used as a precursor of important methylmalonyl-CoA building blocks.</P>
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( Sang Hoon Kwon ),( Jung Soo Eun ),( Young Mo Kang ),( Eon Jeong Nam ) 대한내과학회 2014 대한내과학회 추계학술대회 Vol.2014 No.1
Dermatomyositis (DM) is an idiopathic infiammatory myopathy with characteristic cutaneous manifestations such as heliotrope rash on eyelids and Gottron`s papule. The myopathy in DM shows proximal and often symmetric muscle involvement in the extremities (hip, thigh and shoulder girdle) but rarely affects paraspinal muscles. Here, we report the case of a 43-year old man diagnosed as DM presenting predominant involvement of psoas and paraspinal muscles. Case: A 43-year-old man presented with diffi culty of supine to sitting position and lower back pain for 3 months. On physical exams, he presented scaling, erythematous, itchy cutaneous rashes on dorsum of hands, scalp, and face, especially eyelids. He did not sit on his own in supine position and showed only a low grade muscle weakness (grade 4/5) on lower proximal extremities. Laboratory fi ndings confi rmed the muscle enzyme elevation: CPK 595 U/L (N<170), aldolase 17.8 U/L (N<7.6), serum myoglobin 442 ng/mL (N: 14-106), and LDH 1039 U/L (N: 240-480). Other laboratory fi ndings were ESR 39 mm/hr (N<10), and CRP 1.63 mg/dL (N<0.5). ANA and antibodies were negative. EMG showed polyphasic motor unit potential and increased insertional activity on psoas, paraspinal and gluteus medius muscles. Positron emission tomography demonstrated increased 18FDG uptake in psoas and paraspinal muscles but no 18FDG uptake in proximal muscles of upper and lower extremities. MRI showed hyperintense signals on T2-weighted and gadolinium-enhanced T1-weighted images on psoas and paraspinal muscles. Paraspinal muscle biopsy revealed diffuse infi ammatory endomysial and perivascular infi ltrates, which were compatible with infi ammatory myopathy. Hewas diagnosed as DM with predominant involvement of paraspinal and psoas muscle involvement. He was treated with high dose of steroid and immunosuppressants and showed decreased muscle enzymes and improved muscular weakness.
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The Interpretation of DST Data for Donghae - 1 Gas Field , block VI - 1 , Korea
Sung, Won Mo,Ryou, Sang Soo,Ra, Seung Hun,Kwon, Sun II 한국화학공학회 2001 Korean Journal of Chemical Engineering Vol.18 No.1
Donghae-1 gas field is located in Ulleung basin at offshore Ulsan, Korea, and its recoverable reserve is expected to be 170 to 200 BCF (Billion cubic feet). The held was confirmed to have potential gas and condensate reserves from an exploration well in 1998 and two appraisal wells in 1999. This field consists of five zones, with an average reservoir depth of about 7,000 to 8,000 ft. In this study, we have performed an analysis of Gorae V DST (Drillstem test) #2 for testing B4 zone which has the biggest reserves and Gorae V-1 DST #2 for testing B3 and B4 zones simultaneously among DST data achieved in a total of 11 zones at three wells. The pressure and flow rate recorded from two tested zones were used to obtain the reservoir characteristics and the well productivity. For pressure transient test data, we carried out the analysis of reservoir permeability, skin factor, wellbore storage effect and barrier effect by using the Horner plot and type curve matching methods. Also, with the deliverability test data, we estimated the absolute open flow which is the maximum flow rate of the gas well, and extracted the correlations representing production rate with reservoir pressure. According to the analysis, Gorae V DST #2 of B4 zone has a permeability and skin factor of 37 and (Millidarcy), 4.54, and Gorae V-1 DST #2 of B3 and B4 zones has 23 and and 21.0, respectively. It was also found that the wellbore storage effect was not significant for the two wells tested. From the deliverability test analysis, the AOF (Absolute open flow) of the Gorae V DST #2 is 152.8 MMSCFD (Million standard cubic feet per day), and that of the Gorae V-1 DST #2 is calculated to be 68.2 MMSCFD.
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Kwon, Ou Kwang,Ryou, Sang Soo,Sung, Won Mo 한국화학공학회 2001 Korean Journal of Chemical Engineering Vol.18 No.1
This study presents the development of a four-phase, four-fluid flow pipeline simulator to describe simultaneous flow of gas, oil, water, and hydrate through a pipeline. The model has been equipped with a phase behavior model and hydrate equilibrium model to efficiently estimate thermodynamic and hydrodynamic properties of multicomponent mixtures. The governing equations are formulated for describing the physical phenomena of mass, momentum, and heat transfers between the fluids, and the wall. The equations are solved by utilizing the implicit finitedifference method on the staggered-grid system which can properly describe the boundary conditions as well as phase appearance or disappearance. The developed pipeline simulator has been validated against the field data presented by a previous investigator, and their matches are found to be relatively excellent. The model also has been applied to a multi-component, four-phase flow system in order to examine the transient flow characteristics in pipeline. Also, the potential and the location of hydrate formed in the pipeline have been studied by analyzing the flow characteristics. As a result, it was found that a pipeline system flowing gas, oil, water, and hydrate could be optimized by systematically investigating the hydrodynamic variables for the prevention of hydrate formation.
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Sung Kwon Kim,윤홍인,Wan-Soo Yoon,Jin Mo Cho,Jangsup Moon,Kyung Hwan Kim,김세훈,김영일,김영준,Ho Sung Kim,도윤식,박재성,Ji Eun Park,서영범,Kyoung-Su Sung,송진호,Chan Woo Wee,Se-Hoon Lee,임도훈,Jung Ho Im,장종희,한명훈,홍제범,Kihwan Hwan 대한뇌종양학회 2020 Brain Tumor Research and Treatment Vol.8 No.1
Background The Guideline Working Group of the Korean Society for Neuro-Oncology (KSNO) conducted a nationwide questionnaire survey for diverse queries faced in the treatment of brain tumors. As part II of the survey, the aim of this study is to evaluate the national patterns of clinical practice for patients with diffuse midline glioma and meningioma. Methods A web-based survey was sent to all members of the KSNO by email. The survey included 4 questions of diffuse midline glioma and 6 questions of meningioma (including 2 case scenarios). All questions were developed by consensus of the Guideline Working Group. Results In the survey about diffuse midline glioma, 76% respondents performed histologic confirmation to identify H3K27M mutation on immunohistochemical staining or sequencing methods. For treatment of diffuse midline glioma, respondents preferred concurrent chemoradiotherapy with temozolomide (TMZ) and adjuvant TMZ (63.8%) than radiotherapy alone (34.0%). In the survey about meningioma, respondents prefer wait-and-see policy for the asymptomatic small meningioma without peritumoral edema. However, a greater number of respondents had chosen surgical resection as the first choice for all large size meningiomas without exception, and small size meningiomas with either peritumoral edema or eloquent location. There was no single opinion with major consensus on long-term follow-up plans for asymptomatic meningioma with observation policy. As many as 68.1% of respondents answered that they would not add any adjuvant therapies for World Health Organization grade II meningiomas if the tumor was totally resected including dura. Conclusion The survey demonstrates the prevailing clinical practice patterns for patients with diffuse midline glioma and meningioma among members of the KSNO. This information provides a point of reference for establishing a practical guideline in the management of diffuse midline glioma and meningioma.
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Numerical Predictions of the Load-Displacement Curves of Rock-Socketed Concrete Piles
Kwon, Oh-Sung,Kim, Jeong-Hwan,Jeon, Kyung-Soo,Kim, Myoung-Mo Korean Geotechnical Society 1999 지반 : 한국지반공학회지 Vol.15 No.3
암반 근입말뚝의 설계기준은 일반적으로 침하량 기준을 사용하므로 설계단계에서 암반근입말뚝의 하중-변위$(\sigma-\sigma)$ 거동을 잘 이해해야 하나 그 거동의 복잡성으로 인해 하중-변위 거동을 파악하기가 어렵다. 이를 위해 먼저 현장타설 콘크리트 말뚝에 대해 현장재하시험을 실시하여 그 결과로부터 암반 근입말뚝의 하중-변위 거동을 고찰하였고, 상용 프로그램인 ELAC을 이용하여 하중-변위 곡선의 형상에 큰 영향을 주는 입력변수를 변화시켜 가며 말뚝의 하중-변위 관계를 모사하였다. 최종적으로 수치해석 입력데이터와 현장 지반조사결과의 관계를 일반화함으로써 지반공학자가 설계단계에서 적절한 현장데이터를 이용하여 하중-변위 거동을 예측할 수 있도록 하였다. The settlement limit concept is generally adopted as design criteria of rock-socketed pile foundations, therefore, the load-displacement$(\sigma-\sigma)$ behavior of the rock-socketed piles should be well understood at the design stage, which, however, is hard to achieve due to its complexity. To help this out, field pile load tests are executed on cast-in-situ concrete piles, first, to figure out the $\sigma$-$\delta$ behavior of rock-socketed piles. Next, the $\sigma-\sigma$ relations of the piles are simulated numerically using commercial package program(ELAC) varying a couple of input data which are sensitive in shaping the $\sigma$-$\delta$ curves. Finally, the relation between the best input data for the numerical simulations and the geotechnical field data are cultivated to generalize the numerical simulation procedures, which enables geotechnical engineers to predict the $\sigma$-$\delta$ behavior at the design stage, if appropriate geotechnical field data are provided.
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