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PI4KII activity-dependent Golgi complex targeting of Aplysia phosphodiesterase 4 long-form mutant
전용우,강봉균,장덕진,이진아 한국통합생물학회 2017 Animal cells and systems Vol.21 No.5
The compartmentalization of cAMP by specifically targeted phosphodiesterases (PDEs) contributes to signal regulation in defined regions of cells. We previously demonstrated that the 20 N-terminal amino acids of Aplysia PDE4 (ApPDE4) long-form (L(N20)) and the two mutants of L(N20) were localized to the Golgi complex. However, the molecular mechanisms underlying the Golgi complex targeting of ApPDE4 long-form and its mutated forms are not clear. In the present study, we show that the Golgi complex targeting of L(N20/C14,15S)-enhanced green fluorescent protein (EGFP) was antimycin A-, phenylarsine oxide (PAO)-, and adenosine-sensitive, but insensitive to high concentrations of wortmannin. On the other hand, the Golgi complex targeting of L(N20)-EGFP and L(N20/C3,14S)-EGFP was antimycin A- and PAO-insensitive. These results suggest that the Golgi-localized lipid kinase protein, phosphatidylinositol 4-kinase type II alpha (PI4KIIα), the activity of which is inhibited by PAO and adenosine, but not by high concentrations of wortmannin, is likely involved in the Golgi complex targeting of L(N20/ C14,15S)-EGFP. In addition, subcellular localization of L(N20/C14,15S)-EGFP, but not L(N20)-EGFP or L(N20/C3,14S)-EGFP, was changed from the Golgi complex only to both the endoplasmic reticulum (ER) and the Golgi complex following treatment with a palmitoylation inhibitor, 2-bromo palmitate. Taken together, our results suggest that L(N20/C14,15S)-EGFP, but not L(N20)-EGFP or L(N20/C3,14S)-EGFP, is localized to the Golgi complex in a PI4KII activity- and palmitoylationdependent manner. Therefore, phosphatidylinositol 4-phosphate (PI4P) generated by PI4KIIα at the Golgi complex might play a key role in the Golgi complex targeting of L(N20/C14,15S)-EGFP.
전용우,김선환,백진협,김진아,이용경,민진혜,조형래 대한마취통증의학회 2018 Korean Journal of Anesthesiology Vol.71 No.3
Background: Patients who undergo urinary catheterization may experience postoperative catheter-related bladder discomfort (CRBD). Previous studies have indicated that drugs with antimuscarinic effects could reduce the incidence and severity of CRBD. Accordingly, this study was carried out to investigate whether nefopam, a centrally acting analgesic with concomitant antimuscarinic effect, reduces the incidence and severity of CRBD. Methods: Sixty patients with American Society of Anesthesiologists physical status I and II and aged 18–70 years who were scheduled to undergo elective ureteroscopic litholapaxy participated in this double-blinded study. Patients were divided into control and nefopam groups, comprising 30 patients each. In the nefopam group, 40 mg nefopam in 100 ml of 0.9% saline was administered intravenously. In the control group, only 100 ml of 0.9% saline was administered. All patients had a urethral catheter and ureter stent inserted during surgery. The incidence and severity of CRBD, numerical rating scale (NRS) score of postoperative pain, rescue pethidine dose, and side effects were recorded in the post-anesthesia care unit after surgery. Results: The incidence (P = 0.020) and severity (P < 0.001) of CRBD were significantly different between the control group and the nefopam group. The NRS score of postoperative pain (P = 0.006) and rescue dose of pethidine (P < 0.001) were significantly higher in the control group than in the nefopam group. Conclusions: Intravenous administration of nefopam in patients scheduled to undergo ureteroscopic litholapaxy reduced the incidence and severity of CRBD, NRS score of postoperative pain and analgesic requirements.
전용우,신재용,지종현,조영대,박은철 대한예방의학회 2018 Journal of Preventive Medicine and Public Health Vol.51 No.2
Objectives: Albuminuria has emerged as a biomarker for several medical conditions, and vitamin D has received attention due to its associations with various disorders. We evaluated the association between low serum vitamin D levels and prevalent albuminuria by sex in the Korean general population. Methods: We analyzed 9823 participants (4401 males, 5422 females) from the Korea National Health and Nutrition Examination Survey 2011-2012 (KNHANES V-2), and categorized them as having a normal range of vitamin D levels, vitamin D insufficiency, or vitamin D deficiency. A multivariable logistic regression model was used to compare the risk of albuminuria across these groups. Stratified analyses were conducted by smoking status, obesity, and renal function. Results: Albuminuria was found in 325 of the 4401 male participants (7.4%) and in 455 of the 5422 female participants (8.4%). Among the males, vitamin D deficiency was associated with an odds ratio (OR) for albuminuria of 1.78 (95% confidence interval [CI], 1.07 to 2.97, p<0.05). However, such an association was not found in females. The association was stronger in male current smokers (OR, 3.54; 95% CI, 1.47 to 8.50; p=0.005). Conclusions: The findings of this study suggest that sex differences exist in the association between serum vitamin D deficiency and albuminuria. Additionally, we observed that the association was stronger in current smokers than in the overall male population, but was not seen in non-smokers. Therefore, different approaches by sex and smoking status might be needed when considering using vitamin D as a biomarker for renal function.
Novel GFP-fused protein probes for detecting phosphatidylinositol-4-phosphate in the plasma membrane
전용우,이진아,장덕진 한국통합생물학회 2019 Animal cells and systems Vol.23 No.3
Phosphatidylinositol-4-phosphate (PI4P) plays a crucial role in cellular functions, including protein trafficking, and is mainly located in the cytoplasmic surface of intracellular membranes, which include the trans-Golgi network (TGN) and the plasma membrane. However, many PI4P-binding domains of membrane-associated proteins are localized only to the TGN because of the requirement of a second binding protein such as ADP-ribosylation factor 1 (ARF1) in order to be stably localized to the specific membrane. In this study, we developed new probes that were capable of detecting PI4P at the plasma membrane using the known TGN-targeting PI4P-binding domains. The PI4P-specific binding pleckstrin homology (PH) domain of various proteins including CERT, OSBP, OSH1, and FAPP1 was combined with the N-terminal moderately hydrophobic domain of the short-form of Aplysia phosphodiesterase 4 (S(N30)), which aids in plasma membrane association but cannot alone facilitate this association. As a result, we found that the addition of S(N30) to the N-terminus of the GFP-fused PH domain of OSBP (S(N30)-GFPOSBP- PH), OSH1 (S(N30)-GFP-OSH1-PH), or FAPP1 (S(N30)-GFP-FAPP1-PH) could induce plasma membrane localization, as well as retain TGN localization. The plasma membrane localization of S (N30)-GFP-FAPP1-PH is mediated by PI4P binding only, whereas those of S(N30)-GFP-OSBP-PH and S(N30)-GFP-OSH1-PH are mediated by either PI4P or PI(4,5)P2 binding. Taken together, we developed new probes that detect PI4P at the plasma membrane using a combination of a moderately hydrophobic domain with the known TGN-targeting PI4P-specific binding PH domain.