Preparation and Membrane Distribution of Fluorescent Derivatives of Ceramide

Matsufuji, Takaaki; Kinoshita, Masanao; Matsumori, Nobuaki

doi:10.1021/acs.langmuir.8b03176

Cited by 9 publications

(10 citation statements)

References 43 publications

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“…While the pure SM GUVs formed a homogeneous gel phase (Figure 1a), SM/Cer GUVs underwent phase separation between ceramide-rich and ceramidepoor phases, which were visualized as dark and green domains, respectively (Figure 1b), using a fluorescent-lipid analogue (Bodipy-PC) as a marker of the ceramide-poor phase. 21 This result is in line with previous microscopic observations. 22,23 Interestingly, a similar phase separation was observed for Ceranalogues (Figure 1c−e), indicating that the substitution of the 1-OH group by NHAc, N 3 , or NH 2 does not significantly affect their domain formation in pure water (pH 7).…”

Section: ■ Results and Discussionsupporting

confidence: 93%

“…To compare the domain formability between Cer and its analogues (inclusively termed Cers), we prepared giant unilamellar vesicles (GUVs) composed of palmitoyl-sphingomyelin (SM) and Cers (95:5 and 90:10 mol/mol) binary mixtures and visualized the formation of ceramide-rich phase at 25 °C. While the pure SM GUVs formed a homogeneous gel phase (Figure a), SM/Cer GUVs underwent phase separation between ceramide-rich and ceramide-poor phases, which were visualized as dark and green domains, respectively (Figure b), using a fluorescent-lipid analogue (Bodipy-PC) as a marker of the ceramide-poor phase . This result is in line with previous microscopic observations. , Interestingly, a similar phase separation was observed for Cer-analogues (Figure c–e), indicating that the substitution of the 1-OH group by NHAc, N 3 , or NH 2 does not significantly affect their domain formation in pure water (pH 7).…”

Section: Results and Discussionsupporting

confidence: 89%

“…The contents of the Cers are 5 and 10 mol %, which are directly described in the figure. These samples contain 0.2 mol % Bodipy-PC as a marker for the ceramide-poor phase . Thus, the green and dark regions correspond to ceramide-poor and ceramide-rich phases, respectively.…”

Section: Results and Discussionmentioning

confidence: 99%

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Preparation of Nitrogen Analogues of Ceramide and Studies of Their Aggregation in Sphingomyelin Bilayers

et al. 2021

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Ceramides can regulate biological processes probably through the formation of laterally segregated and highly packed ceramide-rich domains in lipid bilayers. In the course of preparation of its analogues, we found that a hydrogen-bond-competent functional group in the C1 position is necessary to form ceramide-rich domains in lipid bilayers [ Matsufuji Matsufuji Langmuir2018]. Hence, in the present study, we newly synthesized three ceramide analogues: CerN3, CerNH2, and CerNHAc, in which the 1-OH group of ceramide is substituted with a nitrogen functionality. CerNH2 and CerNHAc are capable of forming hydrogen bonds in their headgroups, whereas CerN3 is not. Fluorescent microscopy observation and differential scanning calorimetry analysis disclosed that these ceramide analogues formed ceramide-rich phases in sphingomyelin bilayers, although their thermal stability was slightly inferior to that of normal ceramides. Moreover, wide-angle X-ray diffraction analysis showed that the chain packing structure of ceramide-rich phases of CerNHAc and CerN3 was similar to that of normal ceramide, while the CerNH2-rich phase showed a slightly looser chain packing due to the formation of CerNH3 +. Although the domain formation of CerN3 was unexpected because of the lack of hydrogen-bond capability in the headgroup, it may become a promising tool for investigating the mechanistic link between the ceramide-rich phase and the ceramide-related biological functions owing to its Raman activity and applicability to click chemistry.

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Section: ■ Results and Discussionsupporting

confidence: 93%

Section: Results and Discussionsupporting

confidence: 89%

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Preparation of Nitrogen Analogues of Ceramide and Studies of Their Aggregation in Sphingomyelin Bilayers

et al. 2021

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“…Because saturated sphingolipids increase acyl-chain order in the domains they form, among themselves and together with glycerophospholipids and cholesterol, methods that can detect changes in acyl-chain order are often used to detect the lateral segregation of ordered domains. For bilayer membranes, DSC (49), 2 H NMR (50,51), fluorescence spectroscopy (30,31,35,52,53), and microscopy (54,55) have been successfully used to detect the lateral segregation of ordered domains. The fluorescent probe tPa is excellent for detecting the lateral segregation of sphingolipid-rich domains, which have an increased acyl-chain order when compared to the surrounding fluid glycerophospholipid bilayer.…”

Section: Discussionmentioning

confidence: 99%

Lateral Segregation of Palmitoyl Ceramide-1-Phosphate in Simple and Complex Bilayers

Sazzad

Yasuda

Nyholm

et al. 2019

Biophysical Journal

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Ceramide-1-phosphate is a minor sphingolipid with important functions in cell signaling. In this study, we examined the propensity of palmitoyl ceramide-1-phosphate (Cer-1P) to segregate laterally into ordered domains in different bilayer compositions at 23 and 37 C and compared this with segregation of palmitoyl ceramide (PCer) and palmitoyl sphingomyelin (PSM). The ordered-domain formation in the fluid phosphatidylcholine bilayers was determined using the emission lifetime changes of trans-parinaric acid and from differential scanning calorimetry thermograms. The lateral segregation of Cer-1P was examined when hydrated to bilayers in Tris buffer (50 mM Tris, 140 mM NaCl (pH 7.4)). At this pH, Cer-1P was negatively charged. The lateral segregation propensity of Cer-1P in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) bilayers was intermediate between PCer and PSM. Based on differential scanning calorimetry analysis, we observed that the gel domains formed by Cer-1P in POPC bilayers (POPC:Cer-1P 70:30 by mol) were less stable (melting interval 16-37 C) than the corresponding POPC and PCer gel domains at equal composition (melting interval 20-55 C). The gel-phase melting enthalpy was also much lower in Cer-1P (1.5 kcal/mol) than in the PCer-containing POPC bilayers (9 kcal/mol). Cer-1P appeared to be at least partially miscible with PCer domains in POPC bilayers. Cer-1P domains were stabilized in the presence of PSM (POPC:PSM 85:15), similarly as seen with PCer-rich domains. In bilayers at 37 C, with an approximate outer-leaflet cell membrane composition (sphingomyelin and cholesterol enriched, aminophospholipid poor), Cer-1P segregation did not lead to the formation of ordered domains, at least when compared with PCer segregation. In bilayers with an approximate inner-leaflet composition (sphingomyelin poor, cholesterol and aminophospholipid enriched), Cer-1P also failed to form ordered domains. PCer segregated into ordered domains only after the PCer/cholesterol ratio exceeded an approximate equimolar ratio.

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“…First, he developed a concise method to quantitatively measure the interaction between lipids and MPs (Inada et al 2019b) and revealed that MP-specific lipids have strong effects on the structure and function of MPs (Inada et al 2019a). Second, he is investigating the molecular basis of lipid raft formation by developing new fluorescent derivatives of raft-related lipids (Kinoshita et al 2017;Matsufuji et al 2019). He also explores the actions of drugs and toxins on membranes and/or MPs.…”

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confidence: 99%

Biophysics at Kyushu University

et al. 2020

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It is the largest university in Kyushu and currently has over ten faculties and many institutes. This university has several campuses in Fukuoka city. As shown in Table 1, the annual meeting of the Biophysical Society of Japan has been held five times in Fukuoka city. Kyushu is the westernmost of the four main islands in Japan, and the railway for the bullet train was not extended to Hakata station until 1975. Table 1 includes details of the 10 th annual meeting in 1971 although access was not so good at that time. This history shows that researchers at Kyushu University have been conducting biophysics investigations since the early years of biophysics research in Japan. Kyushu University has many biophysics research groups although it does not have a department of biophysics. Given the interdisciplinary nature of biophysics, these groups belong to their respective faculties and institutes. The Faculty of Science at Ito Campus and the Medical Institute of Bioregulation at Maidashi Campus both have many groups. Eight of these biophysical research groups are introduced in this report. The Medical Institute of Bioregulation was established in 1973. Its aim is to help understand the host-defense mechanisms that operate against infection in humans and other organisms at the molecular, cellular, and individual levels. Daisuke Kohda, the chair of the 46 th Annual Meeting, belongs to this institute (see Table 1). This institute has several biophysical research groups. Here, we introduce the groups of Kohda and Kubota.

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Preparation and Membrane Distribution of Fluorescent Derivatives of Ceramide

Cited by 9 publications

References 43 publications

Preparation of Nitrogen Analogues of Ceramide and Studies of Their Aggregation in Sphingomyelin Bilayers

Preparation of Nitrogen Analogues of Ceramide and Studies of Their Aggregation in Sphingomyelin Bilayers

Lateral Segregation of Palmitoyl Ceramide-1-Phosphate in Simple and Complex Bilayers

Biophysics at Kyushu University

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