• Mass Spectrometry Letters
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• 제13권4호
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• pp.177-183
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• 2022

The monomer and dimer structures of the amyloid fragment Aβ(1-16) sequence formed in H₂O were investigated using electrospray ionization mass spectrometry (MS) and tandem MS (MS/MS). Aβ16 monomers and dimers were indicated by signals representing multiple proton adduct forms, [monomer+zH]ⁿ⁺ (=M^z+, z = charge state) and [dimer+zH]^z+ (=D^z+), in the MS spectrum. Fragment ions of monomers and dimers were observed using collision-induced dissociation MS/MS. Peptide bond dissociation was mostly observed in the D1-D7 and V11-K16 regions of the MS/MS spectra for the monomer (or dimer), regardless of the monomer (or dimer) charge state. Both covalent and non-covalent bond dissociation processes were indicated by the MS/MS results for the dimers. During the non-covalent bond dissociation process, the D³⁺ dimer complex was separated into two components: the M¹⁺ and M²⁺ subunits. During the covalent bond dissociation of the D³⁺ dimer complex, the b and y fragment ions attached to the monomer, (M+b_10-15)^z+ and (M+y_9-15)^z+, were thought to originate from the dissociation of the M²⁺ monomer component of the (M¹⁺+M²⁺) complex. Two different D³⁺ complex geometries exist; two distinguished interaction geometries resulting from interactions between the M¹⁺ monomer and two different regions of M²⁺ (the N-terminus and C-terminus) are proposed. Intricate fragmentation patterns were observed in the MS/MS spectrum of the D⁵⁺ complex. The complicated nature of the MS/MS spectrum is attributable to the coexistence of two D⁵⁺ configurations, (M¹⁺+M⁴⁺) and (M²⁺M³⁺), in the Aβ16 solution.

• Mass Spectrometry Letters
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• 제14권4호
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• pp.153-159
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• 2023

The copper ion, Cu(II), binding sites for amyloid fragment Aβ1-16 (=Aβ16 ) were investigated to explain the biological activity difference in the Aβ16 aggregation process. The [M+Cu+(z-2)H]^z+ (z = 2, 3 and 4, M = Aβ16 monomer) and [D+Cu+(z-2)H]^z+ (z = 3 and 5, D = Aβ16 dimer) structures were investigated using electrospray ionization (ESI) mass spectrometry (MS) and tandem mass spectrometry (MS/MS). Fragment ions of the [M+Cu+(z-2)H]^z+ and [D+Cu+(z-2)H]^z+ complexes were observed using collision-induced dissociation MS/MS. Three different fragmentation patterns (fragment "a", "b", and "y" ion series) were observed in the MS/MS spectrum of the (Aβ16 monomer or dimer-Cu) complex, with the "b" and "y" ion series regularly observed. The "a" ion series was not observed in the MS/MS spectrum of the [M+Cu+2H]⁴⁺ complex. In the non-covalent bond dissociation process, the [D+Cu+3H]⁵⁺ complex separated into three components ([M+Cu+H]³⁺, M³⁺, and M²⁺), and the [M+Cu]²⁺ subunit was not observed. The {M + fragment ion of [M+Cu+H]³⁺} fragmentation pattern was observed during the covalent bond dissociation of the [D+Cu +3H]⁵⁺ complex. The {M + [M+Cu+H]³⁺} complex geometry was assumed to be stable in the [D+Cu+3H]⁵⁺ complex. The {M + fragment ion of [M+Cu]²⁺} fragmentation pattern was also observed in the MS/MS spectrum of the [D+Cu+H]³⁺ complex. The {M + [y₉+Cu]¹⁺} fragment ion was the characteristic fragment ion. The [D+Cu+H]³⁺ and [D+Cu+3H]⁵⁺ complexes were likely to form a monomer-monomer-Cu (M-M-Cu) structure instead of a monomer-Cu-monomer (M-Cu-M) structure.

• 한국결정학회지
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• 제5권2호
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• pp.108-112
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• 1994

C19H20O5, Mr=314.337,삼사정계,PI, a=10.291(2)A, b=11.218(3)A, c=3.059(1)A, α=74.54(2)°,β=1148.84(1)°,r=109.84(2)°,V=883.283(2)A3, λ(Mo Kα)=0.71069A, μ=0.47 mm-1, F(000)=324, 296K, Z=2, Dx=1.18Mgm-3. 1637[F>3σ(F)]개 독립 반사면에 대한 최종 R=0.0580이다. α,β-diphenylsuccinic acid,C16H14O4, 분자들은 O(4)-H˙˙˙O(5)수소결합에 의하여 용체인 acetone과 결합되어 있고 중심대칭관계에 있는 dimer는 분자간의 carboxylic 수소결합 O(1)-H˙˙˙O(2)(-x,-y,-z)에 의하여 결합되어 있다. Dimer간의 가장 가까운 거리 3.288A[O(2)˙˙˙O(2)(-x,-y,-z)]은 분자들의 packing이 van der Waals 력으로 이루어졌음을 보인다.

• Genomics & Informatics
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• 제16권1호
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• pp.2-9
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• 2018

Olfactory receptors (ORs) in mammals are generally considered to function as chemosensors in the olfactory organs of animals. They are membrane proteins that traverse the cytoplasmic membrane seven times and work generally by coupling to heterotrimeric G protein. The OR is a G protein-coupled receptor that binds the guanine nucleotide-binding $G{\alpha}_{olf}$ subunit and the $G{\beta}{\gamma}$ dimer to recognize a wide spectrum of organic compounds in accordance with its cognate ligand. Mammalian ORs were originally identified from the olfactory epithelium of rat. However, it has been recently reported that the expression of ORs is not limited to the olfactory organ. In recent decades, they have been found to be expressed in diverse organs or tissues and even tumors in mammals. In this review, the expression and expected function of olfactory receptors that exist throughout an organism's system are discussed.

• Journal of Microbiology and Biotechnology
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• 제16권11호
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• pp.1832-1836
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• 2006

Mesorhizobium sp. LUK, which utilizes 3-amino-3-phenylpropionic acid as the sole source of nitrogen with high enantioselectivity (E(S)>100), was isolated using enrichment culture. The enzyme involved in the utilization of (S)-3-amino-3-phenylpropionic acid was confirmed to be a transaminase and was purified by 235-folds with a specific activity of 0.72 U/mg. The molecular weight of the purified protein was ca. 47 kDa and the active enzyme was determined as a dimer on gel filtration chromatography. The N-terminal sequence was obtained from the purified protein. Spontaneous decarboxylation of produced ${\beta}-keto$ acids was observed during the chiral resolution of 3-amino-3-phenylpropionic acid.

• Asian-Australasian Journal of Animal Sciences
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• 제17권11호
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• pp.1591-1598
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• 2004

To isolate the $\beta$-galactosidase producing thermophilic bacteria, samples of mud and water were collected from hot springs of avolcanic area near Golden Springs in New Zealand. Among eleven isolated strains, the strain of KNOUC112 produced the highest amounts of $\beta$-galactosidase at 40 h incubation time (0.013 unit). This strain was aerobic, asporogenic bacilli, immobile, gram negative, catalase positive, oxidase positive, and pigment producing. Optimum growth was at 70-72$^{\circ}C$, pH 7.0-7.2, and it could grow in the presence of 3% NaCl. The main fatty acids of cell components were iso-15:0 (30.26%), and iso-17:0 (31.31%). Based on morphological and biochemical properties and fatty acid composition, the strain could be identified as genus Thermus, and finally as Thermus thermophilus by phylogenetic analysis based on 16S rRNA sequence. So the strain is designated as Thermus thermophilus KNOUC112. A gene from Thermus thermophilus KNOUC112 encoding $\beta$-galactosidase was amplified by PCR using redundancy primers prepared based on the structure of $\beta$-galactosidase gene of Thermus sp. A4 and Thermus sp. strain T2, cloned and expressed in E. coli JM109 DE3. The gene of Thermus thermophilus KNOUC112 $\beta$-galactosidase(KNOUC112$\beta$-gal) consisted of a 1,938 bp open reading frame, encoding a protein of 73 kDa that was composed of 645 amino acids. KNOUC112$\beta$-gal was expressed as dimer and trimer in E. coli JM109 (DE3) via pET-5b.

• 미생물학회지
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• 제55권1호
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• pp.9-16
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• 2019

다양한 미생물은 세포와 주변의 과산화를 방지하고 여분의 에너지를 보관하기 위해 ${\alpha}$-acetolactate decarboxylase(ALDC)를 이용해 아세토인을 생성한다. 아세토인은 안전한 식품 향미 개선제이기 때문에 ALDC를 이용한 아세토인 생합성에 많은 산업체가 관심을 가지고 있다. ALDC는 ${\alpha}$-acetolactate의 탈카르복실화 반응을 통해 아세토인을 생산하는 금속 의존 효소이다. 본 논문에서는 고초균 아종 spizizenii의 ALDC(bssALDC) 결정구조를 $1.7{\AA}$ 해상도에서 보고한다. bssALDC는 두 개의 ${\beta}$-sheet가 중앙부를 형성하는 ${\alpha}/{\beta}$ 구조를 가진다. bssALDC는 중앙부의 소수성 상호작용과 주변부의 친수성 상호작용을 통해 이합체를 형성한다. bssALDC는 세 개의 histidine 잔기와 세 개의 물 분자를 이용해 아연 이온에 배위결합한다. 구조와 서열의 비교 분석에 기초하여 아연 이온과 이 주변부 bssALDC 잔기들이 bssALDC의 효소 활성부위임을 제안한다.

• 펄프종이기술
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• 제39권1호
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• pp.16-24
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• 2007

This study was performed to investigate the reaction between alkylketene dimer(AKD) and cellulose molecules in AKD-sized paper sheet. AKD was added to highly beaten($80{\pm}3^{\circ}SR$) SwBKP(ca. 0.8% on pulp) in order to have much AKD retention in the paper sheet. This AKD-sized paper sheet was aged at different temperatures, $60^{\circ}C,\;80^{\circ}C,\;105^{\circ}C\;and\;125^{\circ}C$. Changes in FT-IR spectra of AKD in paper sheet during the ageing were measured. In addition, sizing degrees of the AKD-sized paper sheet pretreated for 30 sec. at $105^{\circ}C$ were measured by HST size tester during the storage at different temperature. IR spectra of AKD-sized paper sheet preheated at $105^{\circ}C$ for 30 sec. showed unchanged spectra two absorption bands at $1849cm^{-1}\;and\;1722cm^{-1}$ which refer to the typical AKD IR bands. However, these typical AKD bands were gradually reduced with increasing ageing, completely disappeared after 6 hrs. and formed new absorption band at $1706cm^{-1}$, which refers to carbonyl stretching vibration of dialkylktone. Eventually the AKD molecule was hydrolyzed to diakylketone without formation of ${\beta}$-ketoester with cellulose in paper sheet. After 6 days ageing, a little amount of ${\beta}$-ketoester bands was identified in 6 or 7 days ageing, because of the absence of water due to long-term heating. The same tendency was observed at different ageing conditions. At the practical papermaking process, AKD reacts prevailing with water, and mostly seems to be hydrolyzed to dialkylketene. Concerned to the sizing development, AKD-sized paper sheet was shown no sizing development at the initial stage of ageing at $60^{\circ}C$ after heating treatment at $105^{\circ}C$ for 30 sec., and gradually increased the sizing degree with increasing ageing, such as Hercules Sizing Tester (HST) 130 see for 12 hr, HST 300 sec. for 3 days and HST 400 sec. for 5 days. It was concluded that hydrolyzed AKD could contributed to the sizing development of the paper sheet.

• Mass Spectrometry Letters
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• 제13권4호
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• pp.166-176
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• 2022

Banana peels are also widely used as bio-adsorbent in the removal of chemicals contaminants and heavy metals from water and soil. GC-MS plays an essential role in the phytochemical analysis and chemo taxonomic studies of medicinal plants containing biologically active components. Intrinsically, with the use of the flame ionization detector and the electron capture detector which have very high sensitivities, Gas chromatography can quantitatively determine materials present at very low concentrations and most important application is in pollution studies. In the present study banana peels were used as bio-adsorbent to remediate the heavy metal contaminated water taken from three different stations located around the industrial belts of Ranipet, Tamilnadu, India. The AAS analysis of the samples shows a decrement of chromium concentration of 98.93%, 96.16% and 96.5% in Station 1, 2 and 3 respectively which proves the efficiency of the powdered peels of Musa paradisiaca. The GC-MS analysis of the control and treated peels of Musa paradisiaca reveals the presence of phytochemicals like Acetic Acid, 1-Methylethyl Ester, DL-Glyceraldehyde Dimer, N-Hexadecanoic Acid, 3-Decyn-2-Ol, 26-Hydroxy, Cholesterol, Ergost-25-Ene-3,5,6,12-Tetrol, (3.Beta.,5.Alpha.,6.Beta.,12.Beta.)-, 1-Methylene-2b-Hydroxymethyl-3, and 3-Dimethyl-4b-(3-Methylbut-2-Enyl)-Cyclohexane in the control banana peels. The banana peels which were used for the treatment reveals the changes and alteration of the phytochemicals. It is concluded that the alteration in phytochemicals of the experimental banana peels were due to adsorption of chromium heavy metal from the sample.

• Toxicological Research
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• 제17권
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• pp.97-104
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• 2001

Three types of proteases (MEF-1, MEF-2 and MEF-3) were purified from the egg cases of Ten-odera sinensis using ammonium sulfate fractionation, gel filtration on Bio-Gel P-60 and affinity chromatography on DEAE Affi-Gel blue gel. The proteases were assessed homogeneous by SDS-polyacrylamide gel electrophoresis and have molecular weight of 31,500, 32,900 and 35,600 Da, respectively. The N-terminal regions of the primary structure were compared and they were found to be different each other. MEFs readily digested the $A\alpha$ - and B$\beta$-chains of fibrinogen and more slowly the ${\gamma}$-chain. The action of the enzymes resulted in extensive hydrolysis of fibrinogen and fibrin, releasing a variety of fibrinopeptides. MEF-1 was inactivated by Cu$^{2+}$ and Zn$^{2+}$ and inhibited by PMSF and chymostatin. MEF-2 was inhibited by PMSF, TLCK. soybean trypsin inhibitor. MEF-3 was only inhibited by PMSF and chymostatin. Antiplasmin was not sensitive to MEF-1 but antithrombin III inhibited the enzymatic activity qf MEF-1. MEF-2 specifically bound to anti plasmin Among the chromogenic protease substrates, the most sensitive one to the hydrolysis of MEFs was benzoyl-Phe-Val-Arg-p-nitroanilide with maximal activity at pH 7.0 and 3$0^{\circ}C$. MEF-1 preferentially cleaved the oxidized B-chain of insulin between Leu15 and Tyr16. In contrast, MEF-2 specifically cleaved the peptide bond between Arg23 and Gly24. D-dimer concentrations increased on incubation of cross-linked fibrin with MEF-1, indicating the enzyme has a strong fibrinolytic activity.ity.