• 제목/요약/키워드: carboxyl terminal

검색결과 96건 처리시간 0.021초

Comparative Biochemical Properties of Proteinases from the Hepatopancreas of Shrimp. -II. Purification of Trypsin from the Hepatopancreas of Penaeus orientalis-

  • Oh Eun-Sil;Kim Doo-Sang;Jung Kyoo-Jin;Pyeun Jae-Hyeung;Heu Min-Soo;Kim Hyeung-Rak
    • Fisheries and Aquatic Sciences
    • /
    • 제1권2호
    • /
    • pp.209-215
    • /
    • 1998
  • Trypsin-like enzyme was purified from shrimp hepatopancreas through Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, and Superdex 75 gel chromatography. Purity of trypsin-like enzyme was increased 69-fold with $44\%$ yield. The enzyme consisted of a single polypeptide chain with a molecular weight (M.W.) of 32 kDa judged by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was completely inactivated by serine enzyme inhibitors such as soybean trypsin inhibitor (SBTI), tosyl-L­lysine chloromethyl ketone (TLCK), and leupeptin. However, the enzyme was not affected by tosyl-L-phenylalanine chloromethyl ketone (TPCK) which is a chymotrypsin specific inhibitor. The enzyme had no activity against benzoyl-tyrosine ethyl ester (BTEE) which is a chymotrypsin specific substrate. The enzyme showed high activity on the carboxyl terminal of Phe, Tyr. Glu, Arg, and Asp. However. no activity was detected against the carboxyl terminal of Pro, Trp, Cys, Gly, Val, and Ala.

  • PDF

α-트로포마이오신의 276 또는 277 아미노산 잔기가 단일 시스테인 잔기로 치환된 돌연변이 트로포마이오신의 액틴친화력 (Actin Affinities of Recombinant α-Tropomyosins That Residues 276 or 277 in the Carboxyl Terminal Region are Individually Substituted to a Cysteine Residue)

  • 김돈규;조영준
    • 생명과학회지
    • /
    • 제19권5호
    • /
    • pp.573-580
    • /
    • 2009
  • 화학적 변형 방식에 의한 트로포마이오신과 액틴의 상호작용을 규명하기 위하여 액틴결합에 중요한 역할을 하는 C-말단부위의 아미노산 잔기 276 또는 277을 단일 시스테인 잔기로 치환한 돌연변이 트로포마이오신을 제조하여 대장균에서 대량 발현시킨 후 액틴 결합력을 측정하였다. 잔기277을 시스테인 잔기로 치환시킨 TM24(QC) 및 TM29(HC)는 액틴 결합 성질을 잃어버렸을 뿐만 아니라 트로포닌 존재 하에서도 액틴결합력이 증가하지 않았다. 이 결과는 잔기 277이 트로포마이신 기능에 중요한 역할을 한다는 것을 제시한다. 반면 잔기 276을 시스테인 잔기로 치환한 TM22(CT) 및 TM23(CA)는 액틴과 비교적 잘 결합하였을 뿐만 아니라 트로포닌 존재 하에서 액틴결합력이 증가하였다. 따라서 TM23(CA)는 시스테인 잔기를 도입하여도 트로포마이오신의 기능을 유지하였으며 향후 화학적 변형 연구를 위한 도구로 중요하게 사용될 수 있을 것이다.

The diverse roles of RNA polymerase II C-terminal domain phosphatase SCP1

  • Harikrishna, Reddy R.;Kim, Hackyoung;Noh, Kwangmo;Kim, Young Jun
    • BMB Reports
    • /
    • 제47권4호
    • /
    • pp.192-196
    • /
    • 2014
  • RNA polymerase II carboxyl-terminal domain (pol II CTD) phosphatases are a newly emerging family of phosphatases that are members of DXDX (T/V). The subfamily includes Small CTD phosphatases, like SCP1, SCP2, SCP3, TIMM50, HSPC129 and UBLCP. Extensive study of SCP1 has elicited the diversified roles of the small C terminal domain phosphatase. The SCP1 plays a vital role in various biological activities, like neuronal gene silencing and preferential Ser5 dephosphorylation, acts as a cardiac hypertrophy inducer with the help of its intronic miRNAs, and has shown a key role in cell cycle regulation. This short review offers an explanation of the mechanism of action of small CTD phosphatases, in different biological activities and metabolic processes.

Synthesis of Various Polymeric Prodrugs of Ibuprofen with PEG and Its Derivative as Polymeric Carriers

  • Lee, Chan-Woo
    • Macromolecular Research
    • /
    • 제12권1호
    • /
    • pp.63-70
    • /
    • 2004
  • We have synthesized various types of poly(ethylene glycol) (PEG)-ibuprofen conjugates by the nucleophilic substitution of bromo-terminated PEG with ibuprofen-Cs salt; PN (Pluronic) was also used in place of PEG. All the bromo-terminated PEGs and PN were obtained in high yield. Conversions of the terminal hydroxyl groups to bromo-termini were quantitative, as were the drug conjugation processes. The Ι$_1$$_3$values obtained from solutions of the ibuprofen-conjugated prodrugs are summarized in relation to those of ibuprofen in water and in aqueous solutions of the original PEG, PN, and several ordinary surfactants. We believe that the fully hydrophilic PEG is completely hydrated and forms no hydrophobic pocket by segment aggregation. These results indicate that the probe environment is significantly hydrophobic, particularly in the solution of prodrug PN, for which the ratio is similar to that obtained from typical micelles of surfactants. The results suggest, therefore, that the present synthetic method is very useful for preparing PEG-based prodrugs from pharmaceuticals having carboxyl functionalities.

평활근 α-트로포마이오신 Gln276잔기의 액틴친화력에 대한 중요성 (Glutamine Residue at 276 of smooth muscle α-tropomyosin is primarily responsible for higher actin affinity)

  • 정선주;조영준
    • 생명과학회지
    • /
    • 제17권2호통권82호
    • /
    • pp.204-210
    • /
    • 2007
  • 평활근 ${\alpha}$-트로포마이오신의 높은 액틴 친화력은 아미노산 잔기 Gln276 및 Thr277에 기인한다는 이전 보고에 따라, 2 잔기 중 어느 잔기가 액틴 친화력에 더 중요한 가를 알아보기 위하여 골격근 트로포마이오신의 His 혹은 Ala 단일 잔기를 각각 Gln 혹은 Thr으로 치환한 돌연변이 트로포마이오신을 제작하여 대장균에서 대량발현 시킨 후 정제하여 액틴 결합력을 측정하였다. 비록 비아세틸화된 트로포마이오신의 경우 Gln 및 Thr 잔기가 최고 액틴친화력을 위해 모두 필요하나, 돌연변이 트로포마이신 중 Gln 잔기를 가진 돌연변이 트로포마이오신들이 다른 돌연변이 트로포마이오신들에 비하여 3에서 4배 높은 액틴친화력을 보였다. 이러한 결과는 평활근 ${\alpha}$-트로포마이오신의 높은 액틴 친화력은 Thr277 잔기보다 Gln276 잔기에 주로 기인한다는 것을 의미한다.

원지와 석창포 혼합추출액의 pCT105로 유도된 신경세포암 세포주에 대한 항치매 효과 (The Effects of anti-Alzheimer in pCT105-induced Neuroblastoma cell lines by Radix Polygalae and Rhizoma Acori Graminei mixture extract)

  • 이성률;강형원;김상태;류영수
    • 동의생리병리학회지
    • /
    • 제17권4호
    • /
    • pp.1037-1049
    • /
    • 2003
  • Numerous lines of evidence indicate that some of the neurotoxicity associated with Alzheimer's disease (AD) is due to proteolytic fragments of the amyloid precursor protein (APP). Most research has focused on the amyloid 6 (M). However, the possible role of other cleaved products of APP is less clear. Lately It has been reported that a recombinant carboxy-terminal 105 amino acid fragment (CT105) of APP induced strong nonselective inward currents in Xenopus oocyte. In a brain with Alzheimer's disease (AD), to investigate the roles of carboxyl-terminal fragment (CT105) of amyloid precursor protein (APP) in apoptosis processes possibly linked to neurodegeneration associated with AD, we examined the effects of the CT of APP with 105 amino acid residues (CT105) on the alteration of apoptosis triggers in neubroblastoma cells. We have investigated whether Radix Polygalae and Rhizoma Acori Graminei mixture extract (RP+RAG) inhibits CT105-induced apoptosis of neuroblastoma cells. We found that RP+RAG inhibits CT105-induced apoptosis in SK-N-SH cells. Treatment of the cells with RP+RAG inhibited CT105-induced DNA fragmentation and Tunel assay of nuclear chromatin and inhibited the caspase-3 expression in SK-N-SH cells. As the result of this study, In RP+RAG group, the apoptosis in the nervous system is inhibited, the repair against the degerneration of neuroblastoma cells by CT105 expression is promoted. These results indicate that RP+RAG possess strong inhibitory effect of apoptosis in the nervous system and repair effect against the degeneration of neuroblastoma cells by CT105 expression

Interleukin-8 수용체와 G$_\alpha$ 16 subunit G protein 간의 분자상호 작용에 관한 연구 (Molecular Interaction Between Interleukin-8 Receptor and G$_\alpha$16 subunit G protein)

  • 하지희;강주섭;고현철;신인철;이창호
    • Biomolecules & Therapeutics
    • /
    • 제8권3호
    • /
    • pp.276-280
    • /
    • 2000
  • In order to identify the domains of the G$_{\alpha}$16 subunit G protein that are responsible for its activation by the Interleukin-8 receptor, a serious of chimeras between G$_{\alpha}$16 and G$_{\alpha}$11 were assessed for their abilities to be activated by these receptors. Co-expression of IL-8 receptor and chimeras in which the carboxyl-terminal regions of G$_{\alpha}$11 were replaced from 30 up to 156 amino acid residues with the corresponding regions of G$_{\alpha}$16 demonstrated that C-terminal 156 amino acid residues of the G$_{\alpha}$16 were not sufficient to confer IL-8 receptor interaction specificity. Testing of a reciprocal serious of chimeras composed of G$_{\alpha}$16 sequences at the amino terminus and G$_{\alpha}$11 sequences at the carboxyl terminals revealed that sequences extending from the amino tar- minus to amino acid 209 of G$_{\alpha}$16 were sufficient to 7ndow the chimera with 75-80% of interaction specificity for 7-8-induced activation. These results suggest th,.7t combined interactions of the C-terminal 30 amino acid residues and certain domains extending from the arts.ino terminus to amino acid 209 of Gal 6 protein may be involved in its couplings to IL-8 receptor.tain domains extending from the arts.ino terminus to amino acid 209 of Gal 6 protein may be involved in its couplings to IL-8 receptor.

  • PDF

COOH-Terminal Animo Acids of Tethered-Buman Glycoprotein Bormone $\alpha$-Subunit Play an Important Role for Secretion

  • Min, K.S;Yoon, J.K.
    • 한국가축번식학회지
    • /
    • 제26권4호
    • /
    • pp.395-399
    • /
    • 2002
  • Human chorionic gonadotropin (hCG) is a member of the glycoprotein hormone family which includes FSH. hCG TSH. These hormone family is characterized by a heterodimeric structure composed a common $\alpha$-subunit noncovalently linked to a hormone specific $\beta$-subunit. To determine u and $\beta$ -subunits can be synthesized as a single polypeptide chain (tethered-hCG) and also display biological activity, the tethered-hCC and -FSH molecule by fusing the carboxyl terminus of the hCG $\beta$-subunit to the amino terminus of the $\alpha$-subunit was constructed. To determine the importance of $\alpha$ COOH -terminal amino acid, we also deleted the $\alpha$ COOH-terminal amino acids. The expressing vectors were transfected into CHO-K 1 cells. The tethered-wthCG and -wtFSH was efficiently secreted. The $\alpha$ Δ83hCG and $\alpha$ Δ 83FSH mutants had no secretion. These results are the first conclusive evidence that COOH-terminal amino acids are very important for secretion in human glycoprotein hormone $\alpha$-subunit. These results demonstrated that the $\alpha$ Δ83hCG and $\alpha$ Δ 83FSH mutants could be play a pivotal role in the secretion of tethered-molecule.

중쇄지방산의 구조적 차이에 따른 병원성 세균에 대한 항균활성 변화 (Effect of structural variation of medium chain fatty acids on antibacterial activities against pathogenic bacteria)

  • 최주현;손수현;김학렬
    • Journal of Applied Biological Chemistry
    • /
    • 제66권
    • /
    • pp.73-80
    • /
    • 2023
  • 일반적으로 여러 다양한 지방산들은 넓은 범위의 미생물들에 대해 항균활성을 나타내는 것으로 알려져 있다. 그러나 지방산의 구조에는 다양한 구조적 차이가 있음에도 불구하고 지방산의 구조적 차이에 따른 항균활성의 변화에 대해서는 많이 알려져 있지 않다. 본 연구는 중쇄지방산들을 대상으로 카복실기의 숫자와 위치에 따른 항균활성의 차이를 잘 알려져 있는 식중독균들을 대상으로 조사하였다. 전반적으로 단일카복실기를 갖는 중쇄 지방산이 이중카복실기를 갖는 중쇄지방산에 비해 더 강한 항균활성을 나타내었지만 두 종류의 중쇄지방산 모두 일반 항생제들과 함께 사용될 경우 매우 높은 시너지 효과를 나타낸다는 사실이 추가적으로 확인되었다.