Symposia / S7-3 : CD38 , a Leukocyte Receptor and Ectoenzyme

김우현(Uh Hyun Kim) 한국생화학분자생물학회 (구 한국생화학회) 1999 생화학분자생물학회 춘계학술발표논문집 Vol.- No.-

Isolation and in vitro translation of the messenger RNA coding for Neurospora crassa NAD glycohydrolase

김우현,서정선,김형로,Kim, Uh-Hyun,Seo, Jeong-Sun,Kim, Hyung-Rho 생화학분자생물학회 1986 한국생화학회지 Vol.19 No.1

Fries minimal 배지와 zinc-nitrogen 결핍 배지에서 각각 배양한 Neurospora crassa로부터 messenger RNA을 얻었다. cell-free reticulocyte system을 이용하여 Neurospora mRNA로부터 in vitro translation 산물을 얻고 면역침전법을 이용하여 이 단백합성 산물내에서 NAD glycohydrolase (EC 3.2.2.5)를 확인하였다. zinc-nitrogen 결핍배지에서의 Neurospora RNA와 mRNA양은 정상배지에서의 양에 비하여 감소하였으나 mRNA의 단백합성능력에는 이상을 보이지 않았다. zinc-nitrogen 결핍배지에서 자란 Neurospora에서 NAD glycohydrolase 특이적 mRNA의 합성을 효소의 비활성도의 증가에 비례하여 증가되지 않은 것으로 보아 NAD glycohydrolase의 증가는 transcription level에서의 조절이 아닌 것으로 사료된다. SDS-polyacrylamide gel 전기영동과 isoelectrofocusing에 의한 단백 합성 산물내의 NAD glycohydrolase 분자량과 pI는 각각 6,000 dalton과 9.0으로서 cell-free reticulocyte system에서의 in vitro translation에서는 post-translational modification이 일어나지 않았음을 시사해 주었다. The in vitro translation products were obtained by translating the isolated Neurospora messenger RNA from Neurospora crassa cultured in Fries minimal medium and zinc and nitrogen deficient medium and the NAD glycohydrolase (NADase) (EC3.2.2.5) was identified in the translation products by the immunoprecipitation methods. The yields of total RNA and messenger RNA from Neurospora crassa grown in zinc and nitrogen deficient media were lower than those from control media but the function of the synthesis of proteins was not affected. The synthesis of NADase-specific mRNA seemed not to be directly proportional to the increase of the specific activity of the enzyme. The increase of NADase in Neurospora crassa grown in zinc and nitrogen deficient seemed not to be ascribed to the regulation in the level of transcription. The molecular weight and the isoelectric point of NADase in the translational products was around 6,000 dalton and pI 9.0, respectively which were different from the other data obtained from purified Neurospora NADase, suggesting that this translation product was not processed post-translational modification.

Neurospora crassa 의 NAD glycohydrolase messenger RNA 정제 및 in vitro translation

김우현,서정선,김형로 ( Uh Hyun Kim,Jeong Sun Seo,Hyung Rho Kim ) 생화학분자생물학회 1986 BMB Reports Vol.19 No.1

The in vitro translation products were obtained by translating the isolated Neurospora messenger RNA from Neurospora crassa cultured in Fries minimal medium and zinc and nitrogen deficient medium and the NAD glycohydrolase (NADase) (EC3.2.2.5) was identified in the translation products by the immunoprecipitation methods. The yields of total RNA and messenger RNA from Neurospora crassa grown in zinc and nitrogen deficient media were lower than those from control media but the function of the synthesis of proteins was not affected. The synthesis of NADase-specific mRNA seemed not to be directly proportional to the increase of the specific activity of the enzyme. The increase of NADase in Neurospora crassa grown in zinc and nitrogen deficient seemed not to be ascribed to the regulation in the level of transcription. The molecular weight and the isoelectric point of NADase in the translational products was around 6,000 dalton and pI 9.0, respectively which were different from the other data obtained from purified Neurospora NADase, suggesting that this translation product was not processed post-translational modification.

Glycosyl - Phosphatidylinositol 막 단백질 연결의 구조와 기능

김우현 ( Uh Hyun Kim ) 생화학분자생물학회 1991 생화학총설집 Vol.3 No.-

백일해 독소에 의한 중성 백혈구 chemotaxis 의 억제에 관한 연구

김정수,김우현,노혜원,김종석,박진우,김형로 ( Jung Soo Kim,Uh Hyun Kim,Hye Won Rho,Jong Suk Kim,Jin Woo Park,Hyung Rho Kim ) 생화학분자생물학회 1989 BMB Reports Vol.22 No.2

It has been suggested that pertussis toxin (PT) inhibits the chemotaxis of neutrophil by modifying signal transduction system coupled to phospholipase C. The PT substrate, probably the inhibitory guanine nucleotide-binding regulatory component of adenylate cyclase (Gi) or an analogous protein, is hence proposed to mediate N-formyl-methionyl-leucyl-phenylalanine (fMLP) receptor-linked activation of the phospholipase C. The aim of this investigation was to determine whether the role played by PT in this system, is dependent upon the ability of the A-protomer of the toxin to ADP-ribosylate Gi or secondary to the binding of the B-oligomer of the toxin to the surface of target cells. PT and methylated PT (mPT) were employed in parallel to assess the involvement of two distinct biologic activities of the toxin in the inhibition of chemotaxis and its related events. Our preparation of mPT retained its ability to ADP-ribosylate Gi, but the pretreatment of mPT to neutrophils did show the inhibition of the chemotaxis as well as the other events related to the phospholipase C, indicating that the involvement of PT is not solely dependent on its capacity to ADP-ribosylate Gi, but rather related to the role of B-oligomer of the toxin.

The Inhibition of Neutrophil Chemotaxis by Pertussis Toxin

김정수,김우현,노혜원,김종석,박진우,김형로,Kim, Jung-Soo,Kim, Uh-Hyun,Rho, Hye-Won,Kim, Jong-Suk,Park, Jin-Woo,Kim, Hyung-Rho 생화학분자생물학회 1989 한국생화학회지 Vol.22 No.2

백일해 독소(PT)는 phospholipase C와 연결된 신호전달 체계를 변형시켜 중성 백혈구의 chemotaxis를 억제한다고 알려져 왔다. fMLP 수용체에 연결되어 phospholipase C가 활성화 되는데는 adenylate cyclase 체계를 억제하는 guanine nucleotide의 결합 조절 단백질(Gi 단백질) 또는 상동성 단백질이 신호전달을 중계할 것이라고 생각되고 있다. 본 연구에서는 이 체계에 대한 PT의 역할이 A-protomer의 ADP-ribosyltransferase 작용에 의한 Gi 단백질의 변형 때문인지 또는 B-oligomer의 세포 표면 결합에 의한 이차적인 효과인지를 구명하였다. PT와 메틸화 PT(mPT)는 chemotaxis 억제 및 관련된 현상들에 서로 다른 생물학적 활성을 가진 독소의 두 부분중 어느 부분이 관여하는지를 알기 위하여 이용하였다. 조제한 mPT는 ADP-ribosyltransferase 활성을 유지하고 있음을 확인하였고 이 mPT는 chemotaxis 억제능력이 없음을 관찰함과 동시에 그에 수반되는 생화학적 사건들에서도 일관성 있는 결과를 얻었기에 PT에 의한 chemotaxis의 억제는 A-protomer보다는 B-oligomer의 역할과 관계되리라 사료된다. It has been suggested that pertussis toxin (PT) inhibits the chemotaxis of neutrophil by modifying signal transduction system coupled to phospholipase C. The PT substrate, probably the inhibitory guanine nucleotide-binding regulatory component of adenylate cyclase (Gi) or an analogous protein, is hence proposed to mediate N-formyl-methionyl-leucyl-phenylalanine (fMLP) receptor-linked activation of the phospholipase C. The aim of this investigation was to determine whether the role played by PT in this system, is dependent upon the ability of the A-protomer of the toxin to ADP-ribosylate Gi or secondary to the binding of the B-oligomer of the toxin to the surface of target cells. PT and methylated PT (mPT) were employed in parallel to assess the involvement of two distinct biologic activities of the toxin in the inhibition of chemotaxis and its related events. Our preparation of mPT retained its ability to ADP-ribosylate Gi, but the pretreatment of mPT to neutrophils did show the inhibition of the chemotaxis as well as the other events related to the phospholipase C, indicating that the involvement of PT is not solely dependent on its capacity to ADP-ribosylate Gi, but rather related to the role of B-oligomer of the toxin.

신장질환에서의 Lactate Dehydrogenase 및 그의 Isoenzyme 활성도 변화

하종영(Chong Y . Ha),박성광(Sung K . Park),강성귀(Sung K . Kang),김우현(Uh . H . Kim) 대한내과학회 1989 대한내과학회지 Vol.37 No.2

N/A The analysis of the 5 different isoenzymes of lactate dehydrogenase(LDH) is of great value in the differential diagnosis of various diseases. In order to investigate the change of LDH isoenzymes in several renal diseases, 44 patients with Korean hemorrhagic fever(KHF), 10 patients with chronic renal failure(CRF), 10 patients with nephrotic syndrome(NS), and 15 healthy subjects were studied, The isoenzymes of LDH were determined by the Helena LDH isoenzyme electrophoresis procedure which is a modification of that of Paston and his associates. Results showed that LDH1 was 22.3±2.8, LDH2 29.4±5.1, LDH3 20.8±4.5, LDH4 9.0±2.7 and LDH5 8.8±3.2 mu/ml in healthy subjects. In patients in the oliguric stage of KHF, LDH1 was 63.4±28.5, LDH2 99.7±40.7, LDH3 107.5±39.0, LDH4 41.9±32.8, and LDH5 37.2±26.3mu/ml, while LDH1 was 23.8±11.7, LDH2 38.9±14.6, LDH3 35.9±18.7, LDH4 13.8±8.0, and LDH5 12.7±7.6mu/ml in non-oliguric patients. In patients with CRF, LDH1 was 33.2±10.8, LDH2 41.9±13.3, LDH3 27.7±8.5, LDH4 12.1±6.2, and LDH5 12.3±5.8 mu/ml. In patients with NS, LDH1 was 25.1±4.3, LDH2 33.5±4.9, LDH3 23.1±6.2, LDH4 8.4±3.7 and LDH5 58.4±3.4 mu/ml. In summary, LDH3 activity was elevated in the oliguric stage of Korean hemorrhagic fever and LDH2 was elevated in chronic renal failure and those correlated with the BUN level.