Quantitative proteomic analysis of gingival crevicular fluids from deciduous and permanent teeth

Yumi, Moriya; Obama, Takashi; Aiuchi, Toshihiro; Sugiyama, Tomomi; Endo, Yumiko; Koide, Yoko; Noguchi, Emiko; Ishizuka, M.; Inoue, Mitsuko; Itabe, Hiroyuki; Yamamoto, Masahide

doi:10.1111/jcpe.12696

Cited by 17 publications

(18 citation statements)

References 45 publications

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“…The tryptic digests were extracted twice with 1% formic acid/ 70% acetonitrile and dried in a SpeedVac evaporator. The extracted peptides were dissolved in 0.1% formic acid and analyzed using a DiNa Nano-LC system (KYA Tech Co., Tokyo, Japan) coupled online with Triple TOF 5600 (Sciex, Framingham, MA, USA), as described previously (40). The oxidized peptides were identified using Pro-teinPilot version 4.5 software (Sciex).…”

Section: Detection Of Oxidatively Modified Amino Acid Residues By Lc-mentioning

confidence: 99%

Circulating oxidized LDL, increased in patients with acute myocardial infarction, is accompanied by heavily modified HDL

Sawada

Obama

Koba

et al. 2020

Journal of Lipid Research

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Oxidized LDL (oxLDL) is a known risk factor for atherogenesis. This study aimed to reveal structural features of oxLDL present in human circulation related to atherosclerosis. When LDL was fractionated on an anion-exchange column, in vivo-oxLDL, detected by the anti-oxidized PC (oxPC) mAb, was recovered in flow-through and electronegative LDL [LDL(−)] fractions. The amount of the electronegative in vivo-oxLDL, namely oxLDL in the LDL(−) fraction, present in patients with acute MI was 3-fold higher than that observed in healthy subjects. Surprisingly, the LDL(−) fraction contained apoA1 in addition to apoB, and HDL-sized particles were observed with transmission electron microscopy. In LDL(−) fractions, acrolein adducts were identified at all lysine residues in apoA1, with only a small number of acrolein-modified residues identified in apoB. The amount of oxPC adducts of apoB was higher in the LDL(−) than in the L1 fraction, as determined using Western blotting. The electronegative in vivo-oxLDL was immunologically purified from the LDL(−) fraction with an anti-oxPC mAb. The majority of PC species were not oxidized, whereas oxPC and lysoPC did not accumulate. Here, we propose that there are two types of in vivo-oxLDL in human circulating plasma and the electronegative in vivo-oxLDL accompanies oxidized HDL.

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Section: Detection Of Oxidatively Modified Amino Acid Residues By Lc-mentioning

confidence: 99%

Circulating oxidized LDL, increased in patients with acute myocardial infarction, is accompanied by heavily modified HDL

Sawada

Obama

Koba

et al. 2020

Journal of Lipid Research

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“…Before collection of GCF, the sampling sites were kept in isolation with cotton wool rolls, dried with air, and the tooth surface was wiped to prevent contamination by pellicle or saliva. Two paper points were inserted at three locations into the gingival sulcus of a single maxillary anterior tooth, after which the six paper points were placed in sterile PBS containing protease inhibitor [15,17]. Samples were frozen in liquid nitrogen and stored at − 80 °C until further use.…”

Section: Gcf Collectionmentioning

confidence: 99%

“…After drying the extract, the sample was dissolved in 30 μL of 0.1% formic acid/2% acetonitrile. The LC-MS/MS analysis was performed as described previously [15].…”

Section: Identification Of Protein By Sds-pagementioning

confidence: 99%

“…Finally, the sample was dissolved in 50 μL of 2% acetonitrile solution before subjecting to LC-MS/MS analysis. The LC-MS/MS analysis was performed as described previously [15].…”

Section: Itraq Labelingmentioning

confidence: 99%

“…GCF contains plasma proteins and inflammatory cells, and its production increases as periodontal disease progresses. Using isobaric tags for relative and absolute quantitation (iTRAQ) technique we previously analyzed relative quantitation of proteins in small amounts of oral samples and observed proteins from plasma and neutrophils are enriched in GCF [15]. A recent paper reported an increase in plasma protein was observed in the pellicle layer on the surface of the incisors of patients with gingivitis [16].…”

Section: Introductionmentioning

confidence: 99%

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Comparison of protein profiles of the pellicle, gingival crevicular fluid, and saliva: possible origin of pellicle proteins

et al. 2020

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Background: The pellicle, the acellular organic material deposited on the surface of tooth enamel, has been thought to be derived from saliva. In this study, protein compositions of the pellicle, gingival crevicular fluid, and saliva collected from healthy adults were compared to elucidate the origin of pellicle proteins. Results: The pellicle, gingival crevicular fluid, and saliva from the parotid gland or mixed gland were collected; subsequently, protein expression in samples from the respective individual was compared by SDS-PAGE and mass spectrometry. Following SDS-PAGE, proteins in the major bands were identified by mass spectrometry. The band pattern of pellicle proteins appeared different from those of gingival crevicular fluid, or saliva samples. Using mass spectrometry, 13 proteins in these samples were identified. The relative abundance of the proteins was quantitatively analyzed using mass spectrometry coupled with stable isotope labeling and by western blot. Cystatin S and α-amylase detected in pellicle were enriched in saliva samples, but not in gingival crevicular fluid, by western blot, and their abundance ratios were high in saliva and low in gingival crevicular fluid when analyzed by stable isotope labeling. Serotransferrin, however, was found only in the pellicle and gingival crevicular fluid by western blot and its abundance ratio was low in saliva. Conclusions: Our study revealed that the gingival crevicular fluid appears to contribute to pellicle formation in addition to saliva.

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Salivary levels of hBDs in children and adolescents with type 1 diabetes mellitus and gingivitis

et al. 2022

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Quantitative proteomic analysis of gingival crevicular fluids from deciduous and permanent teeth

Abstract: Neutrophil-related proteins were enriched in deciduous teeth GCF and immunoglobulins in permanent teeth GCF. This suggests that neutrophil accumulation plays a protective role in innate immunity against bacterial infection in gingival tissue of deciduous teeth.

Cited by 17 publications

References 45 publications

Circulating oxidized LDL, increased in patients with acute myocardial infarction, is accompanied by heavily modified HDL

Circulating oxidized LDL, increased in patients with acute myocardial infarction, is accompanied by heavily modified HDL

Comparison of protein profiles of the pellicle, gingival crevicular fluid, and saliva: possible origin of pellicle proteins

Salivary levels of hBDs in children and adolescents with type 1 diabetes mellitus and gingivitis

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