• Title/Summary/Keyword: N-terminal analysis

Search Result 483, Processing Time 0.049 seconds

Identification of a Functionally Relevant Signal Peptide of Mouse Ficolin A

  • Kwon, Sang-Hoon;Kim, Min-Soo;Kim, Dong-Bum;Lee, Keun-Wook;Choi, Soo-Young;Park, Jin-Seu;Kim, Yeon-Hyang;Lee, Young-Hee;Kwon, Hyung-Joo
    • BMB Reports
    • /
    • v.40 no.4
    • /
    • pp.532-538
    • /
    • 2007
  • Mouse ficolin A is a plasma protein with lectin activity, and plays a role in host defense by binding carbohydrates, especially GlcNAc, on microorganisms. The ficolin A subunit consists of an N-terminal signal peptide, a collagen-like domain, and a C-terminal fibrinogen-like domain. In this study, we show that ficolin A can be synthesized and oligomerized in a cell and secreted into culture medium. We also identify a functionally relevant signal peptide of ficolin A by using MS/MS analysis to determine the N-terminal sequence of secreted ficolin A. When the signal peptide of mouse ficolin A was fused with enhanced green fluorescent protein (EGFP), EGFP was released into HEK 293 cell medium, suggesting that the signal peptide can efficiently direct ficolin A secretion. Moreover, our results suggest that the signal peptide of ficolin A has potential application for the production of useful secretory proteins.

Cloning and Expression of the Aminopeptidase Gene from the Bacillus lichenformis In Bacillus subtilis

  • Kim, Jin-Sook;Lee, In-Soo;Lee, Seung-Won;Lee, Young-Phil;Jung, Chul-Ho;Kim, Hyung-Cheol;Choi, Soon-Yong
    • Journal of Microbiology and Biotechnology
    • /
    • v.12 no.5
    • /
    • pp.773-779
    • /
    • 2002
  • A gene (hap) encoding aminopeptidase from the chromosomal DNA of Bacillus licheniformis was cloned. The gene is 1,347 bp long and encodes a 449 amino acid preproprotein with a major mature region of 401 amino acids (calculated molecular mass 43,241 Da). N-Terminal sequence of the purified protein revealed a potential presence of N-terminal propeptide. The deduced primary amino acid sequence and the mass analysis of the purified protein suggested that a C-terminal peptide YSSVAQ was also cleaved off by a possible endogeneous protease. Tho amino acid sequence displayed 58% identity with that of the aminopeptidase from alkaliphilic Bacillus halodurans. This bacterial enzyme was overexpressed in recombinant Escherichia coli and Bacillus subtilis cells. Clones containing the intact hap gene, including its own promoter and signal sequence, gave rise to the synthesis of extracellular and thrmostable enzyme by B. subtilis transformants. The secreted protein exhibited the same biochemical properties and the similar apparent molecular mass as the B. lichenzyormis original enzyme.

Comparative analysis of AGPase proteins and conserved domains in sweetpotato (Ipomoea batatas (L.) Lam.) and its two wild relatives

  • Nie, Hualin;Kim, Sujung;Kim, Jongbo;Kwon, Suk-Yoon;Kim, Sun-Hyung
    • Journal of Plant Biotechnology
    • /
    • v.49 no.1
    • /
    • pp.39-45
    • /
    • 2022
  • Conserved domains are defined as recurring units in molecular evolution and are commonly used to interpret the molecular function and biochemical structure of proteins. Herein, the ADP-glucose pyrophosphorylase (AGPase) amino acid sequences of three species of the Ipomoea genus [Ipomoea trifida, I. triloba, and I. batatas (L.) Lam. (sweetpotato)] were identified to investigate their physicochemical and biochemical characteristics. The molecular weight, isoelectric point, instability index, and grand average of hyropathy markedly differed among the three species. The aliphatic index values of sweetpotato AGPase proteins were higher in the small subunit than in the large subunit. The AGPase proteins from sweetpotato were found to contain an LbH_G1P_AT_C domain in the C-terminal region and various domains (NTP_transferase, ADP_Glucose_PP, or Glyco_tranf_GTA) in the N-terminal region. Conversely, most of its two relatives (I. trifida and I. triloba) were found to only contain the NTP_transferase domain in the N-terminal region. These findings suggested that these conserved domains were species-specific and related to the subunit types of AGPase proteins. The study may enable research on the AGPase-related specific characteristics of sweetpotatoes that do not exist in the other two species, such as starch metabolism and tuberization mechanism.

3D-QSAR Studies of 3,5-disubstituted Quinolines Inhibitors of c-Jun N-terminal Kinase-3

  • Madhavan, Thirumurthy
    • Journal of Integrative Natural Science
    • /
    • v.4 no.3
    • /
    • pp.216-221
    • /
    • 2011
  • c-Jun N-terminal kinase-3 (JNK-3) has been shown to mediate neuronal apoptosis and make the promising therapeutic target for neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, and other CNS disorders. In order to better understand the structural and chemical features of JNK-3, comparative molecular field analysis (CoMFA) was performed on a series of 3,5-disubstituted quinolines derivatives. The best predictions were obtained CoMFA model ($q^2$=0.707, $r^2$=0.972) and the statistical parameters from the generated 3D-QSAR models were indicated that the data are well fitted and have high predictive ability. The resulting contour map from 3D-QSAR models might be helpful to design novel and more potent JNK3 derivatives.

Protein-Protein Interaction between Poly(A) Polymerase and Cyclophilin A in Chemotactic Cells

  • Choi, Hyun-Sook;Kim, Hana;Lee, Changgook;Kim, Youngmi;Lee, Younghoon
    • Bulletin of the Korean Chemical Society
    • /
    • v.35 no.1
    • /
    • pp.83-86
    • /
    • 2014
  • Poly(A) polymerase (PAP) play an essential role for maturation of mRNA by adding the adenylate residues at the 3' end. PAP functions are regulated through protein-protein interaction at its C-terminal region. In this study, cyclophilin A (CypA), a member of the peptidyl-prolyl cis-trans isomerase family, was identified as a partner protein interacting with the C-terminal region PAP. The interaction between PAP and CypA was inhibited by the immunosuppressive drug cyclosporine A. Deletion analysis revealed that the N-terminal 56 residues of CypA are sufficient for the interaction with PAP. Interestingly, we observed that PAP and CypA colocalize in the nucleus during SDF-1-induced chemotaxis, implying that CypA could be involved in the regulation of polyadenylation by PAP in the chemotactic cells.

Analysis of Erythropoietin Glycoform Produced by Recombinant CHO Cells Using the Lectin-Blotting Technique

  • Chang, Kern-Hee;Kim, Kyung-Soo;Kim, Jung-Hoe
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.3 no.1
    • /
    • pp.40-43
    • /
    • 1998
  • The glycosylation pattern of Erythropoietin (EPO), produced by recombinant CHO cells, was studied using the simple and rapid technique of 'Lectin-blotting'. In this experiment we used three different kinds of lectins, MAA(Maackia amurensis agglutinine), RCA(Ricinus communis agglutinine), and DSA(Datura stramonium agglutinine), which bind to the terminal sialic acid, galactose, and the N-acetyllactosamine chain respectively. The lectin-blotting technique was used to analyze the carbohydrate structure of EPO produced in the presence of two physiologically active chemical compounds, ammonium and chloroquine. The effect of the ammonium ion on the glycosylation of EPO was studied because it accumulated in the medium mainly as a by-product of glutamine matabolism. Ammonium chloride significantly inhibited the sialylation of the terminal galactose residue at concentrations of 8mM or more. Chloroquine, a potent inhibitor of glycosylation, inhibited terminal sialylation at concentrations of 100 and 200 $\mu$M, and at a concentration of 300 $\mu$M, also inhibited Nacetyllactosamine chain synthesis.

  • PDF

A Study of Nurses Burden and Attitude on Terminal Cancer Patients (말기암 환자에 대한 간호사의 부담감과 태도 연구)

  • Kim, Yun-Hee
    • Asian Oncology Nursing
    • /
    • v.1 no.1
    • /
    • pp.65-74
    • /
    • 2001
  • This study was to investigate the degree of the nurses' burden and the attitude on the terminal cancer patients, as well as the relationship between two variables using questionnaire. The non-randomized convenient samples were 252 nurses with the experiences in caring the terminal cancer patients more than 1year in 5 university hospitals in Seoul and Inchon city. The cross-sectional one time survey was conduced by using the modified questionnaires on the burden and the attitude on the terminal cancer patients at October, 2000. n SPSS for Window, the demographic information and the degree of the burden and the attitude of subjects were analyzed with descriptive statistics. Pearson correlation coefficiency was used to investigate the relationship between the degree of the burden and the attitude from subjects. The additional analysis were performed to examine the differences the degree of the burden and the attitude by the general characteristics of the nurses using t-test and ANOVA. The result was as follow: 1) The degree of the nurses' burden on the terminal cancer patients was the mean of 2.91 ranged from 2.08 to 3.96. 2) The degree of the nurses' attitude on the terminal cancer patients was the mean of 3.52 ranged from 1.83 to 4.68. 3) There was no significant relationship between the degree of the burden and the attitude on the terminal cancer patients(r=.08, p=.23). However, the burden and. the nursing environment among 4 aspects of the attitude showed a significantly positive relationship each other (r=.16, p=.01). 4) The degree of the nurses' burden was different by the nursing specialties (F=2.79, p=.03) and the professional perspectives on nursing(F=3.52, p=.02). 5) The degree of the nurses' attitude was different by the age(F=5.33, p=.01), the married status(t=3.93, p=.05), nursing specialties (F=7.42, p=.00), the amount clinical experience(F=2.85, p=.04), the job satisfaction (F=10.58, p=.00) and, the professional perspectives on nursing (F=6.30, p=.01).

  • PDF

Factors Associated with Place of Death in Korean Patients with Terminal Cancer

  • Hyun, Min Kyung;Jung, Kyung Hae;Yun, Young Ho;Kim, Young Ae;Lee, Woo Jin;Do, Young Rok;Lee, Keun Seok;Heo, Dae Seog;Choi, Jong Soo;Kim, Sam Yong;Kim, Heung Tae;Hong, Seok-Won
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.12
    • /
    • pp.7309-7314
    • /
    • 2013
  • Aim: To investigate factors that affect the place of death (POD) of terminal cancer patients. Materials and Methods: We recruited 702 consecutive patients (${\geq}18$ years) from 12 centers during July 2005 to October 2006, and 481 completed the questionnaire. In April 2011, we linked the data for 96.0% (n=462) of the deceased patients to the POD using the 2005-2009 death certificate data of Korea's National Statistical Office. The primary outcome variable was POD, and the predictive value of variables pertaining to patients and caregivers was evaluated using univariate and multivariate analyses. Results: Most patients died in a hospital (91.5%, n=441) and age, education, preference for place of terminal care, wish to use hospice/palliative care services, terminal cancer awareness, time between diagnosis and death, and global quality-of-life subscale of the EORTC QLQ-C30 of patients, and education and preference for place of terminal care of caregivers were significant predictors in univariate analyses. On multivariate analysis, patients and caregivers who preferred hospital/palliative care as the terminal care option over home care [adjusted odds ratio (aOR), 2.68; 95% confidential interval (CI), 1.18-7.04 and aOR: 2.65; 95%CI: 1.15-6.09 for patient and caregiver preferences, respectively] and caregivers who were highly educated (aOR, 3.19; 95%CI, 1.44-7.06) were predictors of POD. Conclusions: Most of the terminal cancer patients died in a hospital. Our findings indicate that major predictors of hospital deaths are preference of both the patient and caregiver for hospital/palliative care as the terminal care option and higher education of the caregiver.

Purification and Characterization of Vitellin from the Firefly, Pyrocoelia rufa

  • Kim, Seong-Ryul;Jin, Byung-Rae;Yang, Won-Jin;Kim, Jong-Gill;Kim, Keun-Young;Lee, Sang-Mong;Moon, Byung-Ju;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • v.2 no.2
    • /
    • pp.167-172
    • /
    • 2001
  • The vitellin of firefly, Pyrocoelia rufa, is composed of three polypeptides, designated Vn1 (175 kDa), Vn2 (160 kDa) and Vn3 (45 kDa) in SDS-polyacrylamide gel electrophoresis. Three subunits of vitellin were presented in the female adult hemolymph, ovary and egg extracts, but not observed in the male. This vitellin was purified from the eggs of P. rufa by the FPLC techniques, anion exchange chromatography and gel permeation chromatography. In nature, vitellin of P. rufa has molecular weight of 400 kDa. Western blot analysis using polyclonal antiserum against purified vitellin showed that the antiserum was reacted with the three polypeptides, Vnl, Vn2 and Vn3 from the female adult hemolymph, ovary and egg extracts. Amino acid residues at N-terminus of three subunits were sequenced. The N-terminal sequences of large subunits, Vnl and Vn2, were similar to each other, But, the N-terminal sequences of small subunits Vn3, did not have any signnificant homology with large subunits.

  • PDF

Fragmentation Analysis of rIAPP Monomer, Dimer, and [MrIAPP + MhIAPP]5+ Using Collision-Induced Dissociation with Electrospray Ionization Mass Spectrometry

  • Kim, Jeongmo;Kim, Ho-Tae
    • Mass Spectrometry Letters
    • /
    • v.12 no.4
    • /
    • pp.179-185
    • /
    • 2021
  • Collision-induced dissociation (CID) combined with electrospray ionization mass spectrometry (ESI-MS) was used to obtain structural information on rat islet amyloid polypeptide (rIAPP) monomers (M) and dimers (D) observed in the multiply charged state in the MS spectrum. MS/MS analysis indicated that the rIAPP monomers adopt distinct structures depending on the molecular ion charge state. Peptide bond dissociation between L27 and P28 was observed in the MS/MS spectra of rIAPP monomers, regardless of the monomer molecular ion charge state. MS/MS analysis of the dimers indicated that D5+ comprised M2+ and M3+ subunits, and that the peptide bond dissociation process between the L27 and P28 residues of the monomer subunit was also maintained. The observation of (M+ b27)4+ and (M+ y10)3+ fragment ions were deduced to originate from the two different D5+ complex geometries, the N-terminal and C-terminal interaction geometries, respectively. The fragmentation pattern of the [MrIAPP + MhIAPP]5+ MS/MS spectrum showed that the interaction occurred between the two N-terminal regions of MrIAPP and MhIAPP in the heterogeneous dimer (hetero-dimer) D5+ structure.