Tackykinin Peptide

신송엽,무네카타 에이수케 한국생화학분자생물학회 1992 생화학분자생물학회 소식 Vol.12 No.4

Three mammalian neuropeptide, SP, NKA and NKB, belong to a family of peptides known as tachykinins which share the common sequence Phe-X-Gly-Leu-Met-NH₂. These novel peptides possess a commom spectrum of biological activities including a sensory transmission in nervous system and contractions/relaxation of peripheral smooth muscles. This review will describe recent knowledges and trends on the biological activity, biosynthesis. release, distribution. receptor and antagonist of these tachykinin peptides.

Prokaryotic Selectivity, Bactericidal Mechanism and LPS-neutralizing Activity of Lys-linked Dimeric Peptide of Indolicidin C-terminal Hexapeptide

신송엽 대한화학회 2013 Bulletin of the Korean Chemical Society Vol.34 No.7

Neurokinin군 펩티드유사체들의 모르모트 기관 평활근 수축활성

신송엽,하종명,하배진,장태식,강신원,무나카타 에이스케,Shin, Song-Yub,Ha, Jong-Myung,Ha, Bae-Jin,Jang, Tae-Sik,Kang, Shin-Won,Munekata, Eisuke 생화학분자생물학회 1994 한국생화학회지 Vol.27 No.5

Substance P(SP), Neurokinin A(NKA) 및 Neurokinin B(NKB)는 포유류 유래의 신경펩티드(neuropeptide)로서 Neurokinin군으로 분류되며, Tachykinin족에 속한다. SP, NKA 및 NKB의 생물활성의 차이를 화학구조의 면에서 해명하기 위하여 Neurokinin군 펩티드들의 아미노산 배열을 서로 바꾼 유사체들을 Boc-HF에 의한 고상법에 의하여 화학합성하고, 이 펩티드들의 생물활성을 모르모트 기관(Guinea Pig Trachea : GPT)의 평활근에 대한 근육수축활성을 지표로하여 측정하였다. 본 연구에서는, GPT에 대한 Neurokinin군 펩티드들의 평활근의 수축활성은 카르복실 말단측의 7개의 아미노산이 필수적임을 확인하였다. 또한, 제4위의 아스파르트산, 제7위의 발린이 활성발현에 중요하며, NKA와 NKB의 생물활성의 차이는 제5위의 아미노산(NKA; Ser, NKB; Phe)의 구조적인 차이에 의존함이 시사되었다. Neurokinin peptides (Substance P; SP, Neurokinin A; NKA and Neurokinin B; NKB) are a neuropeptide belong to the mammalian tachykinin family. To elucidate the difference in biological properties among neurokinin peptides from chemical structure, the peptide derivatives of neurokinins were synthesized by the solution and solid-phase method and their contractile activities were measured by contraction assay with the smooth muscle of guinea pig trachea (GPT). This study demonstrated that the C-terminal heptapeptide containing two amino acid residues, $Asp^4$ and $Val^7$, is indispensible for contracting activity in GPT. Further, structureactivity studies indicate that the amino acid residues (Ser and Phe) at position 5 from N-terminus play an important role in showing the difference of biological actvity between NKA and NKB in GPT assay.

Human Epidermal Growth Factor 유사체들의 고상합성과 생물활성

신송엽,하종명 ( Song Yub Shin,Jong Myung Ha ) 생화학분자생물학회 1993 BMB Reports Vol.26 No.3

Human epidermal growth factor (h-EGF) is 53 amino acid polypeptide that stimulates proliferation and differentiation of various cells. To identify critical residues that govern biological activity of h-EGF, six analogues having specific amino acid substitution at 13, 15, 37 and 41 were synthesized by stepwise solid phase method using the Boc-HF strategy. Homogenity and purity of the synthesized peptides were performed by RP-HPLC, FAB-MS, amino acid analysis and thermolytic digestion. The biological activity of these h-EGF analogues was measured by mitogenic stimulation in NIH/3T3 cells. The substitution of aromatic amino acid (Phe) at position 37 and 13 had little effect on mitogenic activity of h-EGF. In contract, the replacement of Ala at these positions occurred a big loss of acitivity (5×10^(-2) and 10^(-3) -fold). This result indicate that aromatic side chain at position 13 and 37 play an important role in biological activity of h-EGF. The semiconservative substitution of Ala for Leu at position 15 and conservative change of Lys for Arg at position 41 also drastically reduced mitogenic activity (10^(-4) and 10^(-5) `-fold). Thus, the hydrophobic bulky group at postion 15 and the guanidino group at position 41 were essential for the biological activity of h-EGF.

Neurokinin 군 펩티드유사체들의 모르모트 기관 평활근 수축활성

신송엽,하종명,하배진,장태식,강신원,종상 영보 ( Song Yub Shin,Jong Myung Ha,Bae Jin Ha,Tae Sik Jang,Shin Won Kang,Eisuke Munekata ) 생화학분자생물학회 1994 BMB Reports Vol.27 No.5

Neurokinin peptides (Substance P; SP, Neurokinin A; NKA and Neurokinin B; NKB) are a neuropeptide belong to the mammalian tachykinin family. To elucidate the difference in biological properties among neurokinin peptides from chemical structure, the peptide derivatives of neurokinins were synthesized by the solution and solid-phase method and their contractile activities were measured by contraction assay with the smooth muscle of guinea pig trachea (GPT). This study demonstrated that the C-terminal heptapeptide containing two amino acid residues, Asp⁴ and Val^7, is indispensible for contracting activity in GPT. Further, structureactivity studies indicate that the amino acid residues (Ser and Phe) at position 5 from N-terminus play an important role in showing the difference of biological activity between NKA and NKB in GPT assay.

Neurokinin A의 구조 및 활성에 관한 연구

신송엽,하종명,하배진,Shin, Song-Yub,Ha, Jong-Myung,Ha, Bae-Jin 생화학분자생물학회 1993 한국생화학회지 Vol.26 No.8

Neurokinin A(NKA)와 Neurokinin B(NKB)는 Tachykinin족 펩티드에 속하는 포유류 유래의 신경펩티드이다. NKA와 NKB의 생물활성의 차이를 화학구조의 면에서 해명하기 위하여 Neurokinin펩티드의 유사체들을 화학합성하고 이 유사체들의 생물활성을 rabbit pulmonary artery(RPA)의 평활근에 대한 근육수축활성을 지표로하여 측정하였다. NKA의 아미노 말단으로부터 제5위의 세린을 페닐알라닌으로 치환시키면 약 1/5의 활성을 보였고, NKB의 아미노 말단으로부터 제5위의 페닐알라닌을 세린으로 치환시키면 NKA로서의 활성이 상승하였다. 이 결과로부터, NKA의 생물활성발현에는 아미노말단으로부터 제5위의 아미노산인 세린과 세린을 포함하는 message segment가 중요한 역할을 담당하고 있음이 시사되었다. Neurokinin A (NKA) is neuropeptide belong to mammalian tachykinin family. To investigate the difference in biological properties between NKA and NKB from the chemical structure, the peptide derivatives of neurokinins were synthesized by solution phase method and their contractile activities were measured by contraction assay with the smooth muscle of rabbit pulmonary artery (RPA). The substituion of Ser for Phe at position 5 in NKA drastically reduced contractile activity. The replacement of Phe at 5 position in NKB with Ser showed increase in contractile activity. These results suggest that message segment including Ser at position 5 has a critical role in the biological activity of NKA.

Neurokinin A 의 구조 및 활성에 관한 연구

신송엽,하종명,하배진,Munekata Eisuke ( Song Yub Shin,Jong Myung Ha,Bae Jin Ha,Munekata Eisuke ) 생화학분자생물학회 1993 BMB Reports Vol.26 No.8

Neurokinin A (NKA) is neuropeptide belong to mammalian tachykinin family. To investigate the difference in biological properties between NKA and NKB from the chemical structure, the peptide derivatives of neurokinins were synthesized by solution phase method and their contractile activities were measured by contraction assay with the smooth muscle of rabbit pulmonary artery (RPA). The substituion of Ser for Phe at position 5 in NKA drastically reduced contractile activity. The replacement of Phe at 5 position in NKB with Ser showed increase in contractile activity. These results suggest that message segment including Ser at position 5 has a critical role in the biological activity of NKA.

Effect of Double Replacement of L-Pro, D-Pro, D-Leu or Nleu in Hydrophobic Face of Amphipathic α-Helical Model Antimicrobial Peptide on Structure, Cell Selectivity and Mechanism of Action

신송엽 대한화학회 2014 Bulletin of the Korean Chemical Society Vol.35 No.11

In order to investigate the effects of the double replacement of L-Pro, D-Pro, D-Leu or Nleu (the peptoid residue for Leu) in the hydrophobic face (positions 9 and 13) of amphipathic α-helical non-cell-selective antimicrobial peptide L8K9W1 on the structure, cell selectivity and mechanism of action, we synthesized a series of L8K9W1 analogs with double replacement of L-Pro, D-Pro, D-Leu or Nleu in the hydrophobic face of L8K9W1. In this study, we have confirmed that the double replacement of L-Pro, D-Pro, or Nleu in the hydrophobic face of L8K9W1 let to a great increase in the selectivity toward bacterial cells and a complete destruction of α-helical structure. Interestingly, L8K9W1-L-Pro, L8K9W1-D-Pro and L8K9W1-Nleu preferentially interacted with negatively charged phospholipids, but unlike L8K9W1 and L8K9W1-D-Leu, they did not disrupt the integrity of lipid bilayers and depolarize the bacterial cytoplasmic membrane. These results suggested that the mode of action of L8K9W1-L-Pro, L8K9W1-D-Pro and L8K9W1-Nleu involves the intracellular target other than the bacterial membrane. In particular, L8K9W1-L-Pro, L8K9W1-D-Pro and L8K9W1-Nleu had powerful antimicrobial activity (MIC range, 1 to 4 μM) against methicillin-resistant Staphylococcus aureus (MRSA) and multidrug-resistant Pseudomonas aeruginosa (MDRPA). Taken together, our results suggested that L8K9W1- L-Pro, L8K9W1-D-Pro and L8K9W1-Nleu with great cell selectivity may be promising candidates for novel therapeutic agents, complementing conventional antibiotic therapies to combat pathogenic microorganisms.

Structure-Activity Study of Guinea Pig Trachea Tachykinin NK-2 Receptor: Effect of Substitution at the Seventh Position from C-Terminus of Neurokinin A

신송엽,방정규,박장수,강신원,Sin, Song Yeop,Bang, Jeong Gyu,Park, Jang Su,Gang, Sin Won Korean Chemical Society 2000 Bulletin of the Korean Chemical Society Vol.21 No.9

Human Epidermal Growth Factor 유사체들의 고상합성과 생물활성

신송엽,하종명,Shin, Song-Yub,Ha, Jong-Myung 생화학분자생물학회 1993 한국생화학회지 Vol.26 No.3

Human Epidermal Growth Factor(h-EGF)는 세포의 증식과 분화를 촉진시키는 53개의 아미노산으로 이루어진 폴리펩티드이다. h-EGF의 생물활성의 발현에 중요한 잔기를 확인하기 위하여, h-EGF의 제 13, 15, 37 및 41위의 아미노산을 다른 아미노산으로 치환시킨 펩티드들을 Boc-HF strategy를 이용한 단계적 고상합성에 의하여 화학합성하였다. 모든 합성펩티드들의 순도는 역상 고속액체크로마토그래피(RP-HPLC), FAB-mass spectroscopy, 아미노산 분석 및 Thermolysin에 의한 효소분해법 등으로 확인하였고, NIH/3T3세포의 세포증식효과에 의하여 모든 합성펩티드들에 대한 생물학적 활성을 측정하였다. 그 결과, 제 13위와 37위의 티로신을 같은 방향족 아미노산인 페닐알라닌으로 치환시킨 경우 활성의 변화는 거의 없었으나, 같은 부위를 알라닌으로 치환시키면 각각 약 $5{\times}10^{-2}$배 및 $10^{-3}$배로 활성이 감소하였다. 이와 같은 결과는 h-EGF의 생물활성의 발현에는 제 13위와 37위의 방향족 곁사슬이 중요한 역할을 담당하고 있다는 것을 시사한다. 또한, 제 15위의 로이신을 알라닌으로 치환했을 때와 제 41위의 아르기닌을 리진으로 치환했을 때 각각 약 $10^{-4}$배 및 $10^{-5}$배의 심한 활성의 저하를 나타내었다. 따라서 제 15위의 hydrophobic bulky group과 제 41위의 guanidino group은 h-EGF의 기능의 발현에 필수적임을 시사한다. Human epidermal growth factor (h-EGF) is 53 amino acid polypeptide that stimulates proliferation and differentiation of various cells. To identify critical residues that govern biological activity of h-EGF, six analogues having specific amino acid substitution at 13, 15, 37 and 41 were synthesized by stepwise solid phase method using the Boc-HF strategy. Homogenity and purity of the synthesized peptides were performed by RP-HPLC, FAB-MS, amino acid analysis and thermolytic digestion. The biological activity of these h-EGF analogues was measured by mitogenic stimulation in NIH/3T3 cells. The substitution of aromatic amino acid (Phe) at position 37 and 13 had little effect on mitogenic activity of h-EGF. In contract, the replacement of Ala at these positions occurred a big loss of acitivity $(5{\times}10^{-2} \; and \; 10^{-3}-fold)$. This result indicate that aromatic side chain at position 13 and 37 play an important role in biological activity of h-EGF. The semiconservative substitution of Ala for Leu at position 15 and conservative change of Lys for Arg at position 41 also drastically reduced mitogenic activity $(10^{-4} \; and \; 10^{-5}-fold)$. Thus, the hydrophobic bulky group at postion 15 and the guanidino group at position 41 were essential for the biological activity of h-EGF.