Recombinant preparation and characterization of interactions for a calmodulin-binding chromogranin A peptide and calmodulin

Kang, Sunmi,Kang, Jinho,Yoo, Seung Hyun,Park, Sunghyouk JOHN WILEY & SONS LTD 2007 Journal of Peptide Science Vol.13 No.4

<P>Chromogranin-derived peptides have important and varied biological activities. They affect a wide spectrum of targets such as fungal membranes, blood vessels, myocardial cells, and pancreatic cells. Despite the biological significance and the diverse activities, the molecular mechanisms of the interactions between the peptides and the target proteins have not been well understood. Here, we studied the interaction between a chromogranin A–derived peptide (CGA40–65) and its target protein, calmodulin, with NMR spectroscopy. Calmodulin was easily prepared with standard recombinant technology, but CGA40–65 posed challenges requiring multistep procedures. The recombinantly produced peptide retained the calmodulin-binding property of the full-length CGA, as shown by the HSQC binding experiment. By applying resonance assignments, we identified the residues in calmodulin involved in the CGA40–65 binding. We also found that the peak changes are close to those exhibited by the peptides having the wrap-around binding mechanism. Further analysis revealed that the CGA40–65-induced changes are more similar to those by CaMKIp peptide than those by smMLCKp peptide among the wrap-around binding peptides, suggesting that CGA40-65 can be categorized as a CaMKIp-like peptide. Our report is the first residue-resolution mechanistic study involving chromogranin peptides and their target proteins. Our approaches should be applicable to interaction studies involving other chromogranin-derived peptides and their cellular target proteins. Copyright © 2007 European Peptide Society and John Wiley & Sons, Ltd.</P>

근치적 전립샘적출술을 시행한 699예의 전립샘암종에서 신경내분비 분화가 예후에 미치는 영향

김태훈,신은,김백희,최기영 대한병리학회 2008 Journal of Pathology and Translational Medicine Vol.42 No.6

Background : Neuroendocrine differentiation of prostatic carcinoma is known to be associated with a poor prognosis, tumor progression and androgen-independency, and there is currently no successful therapy for this type of tumor. The purpose of this study is to evaluate the prognostic implications of neuroendocrine differentiation in prostatic carcinoma in Korean men. Methods : Six hundreds and ninety nine consecutive cases of radical prostatectomy specimens were systematically processed for topographic mapping. Neuroendocrine differentiation was detected by immunohistochemistry by using antibody to chromogranin. We analyzed the relationship between neuroendocrine differentiation and the clinicopathological prognostic factors, as well as biochemical failure. The neuroendocrine differentiation was evaluated according to the presence of chromogranin-positive cells, the pattern of neuroendocrine cells and the number of neuroendocrine cells, respectively. Results : Neuroendocrine differentiation was detected in 150 out of 699 cases (21.5%). The presence of neuroendocrine differentiation as well as the pattern of neuroendocrine cells was correlated with biochemical failure and the other clinicopathological prognostic factors such as the Gleason score, the pathologic stage, the tumor volume, angiolymphatic invasion, perineural invasion, and the Ki-67 proliferative index (p<0.05). Conclusions : We suggest that neuroendocrine differentiation of prostatic carcinoma is a prognostic factor even in radical prostatectomy specimens for localized prostate cancer. Evaluation of the presence of neuroendocrine differentiation as well as the pattern of neuroendocrine cells is recommended in radical prostatectomy specimens.

Changes of Gastrointestinal Chromogranin-Immunoreactive Cells after Implantation of Colon-26 Murine Carcinoma Cells in BALB/c Mouse

Taesu Ham,Hyeungsik Lee,Jaehyun Lee,Saekwang Ku 한국실험동물학회 2005 Laboratory Animal Research Vol.21 No.3

The changes on the regional distribution and frequency of chromogranin-immunoreactive (CG-IR) cells in gastrointestinal (GI) tract of BALB/c mouse after implantation of murine carcinoma cells, colon-26 were studied by immunohistochemical methods. The experimental animals were divided into two groups; non¬implanted (Sham) and colon-26 (CT-26) cell implanted (Colon-26) groups. Samples were collected from each part of GI tract (fundus, pylorus, duodenum, jejunum, ileum, cecum, colon and rectum) at 28th days after subcutaneously implantation of CT-26 cells (1×10⁵ cell/mouse). CG-IR cells were distributed throughout the whole GI tract except for the rectum in Sham with various frequencies. However, they were restricted to the fundus, pylorus, duodenum and jejunum of Colon-26. A significantly (p<0.01) decrease of CG-IR cells was detected in Colon-26 compared to that of Sham in both fundus and pylorus, but similar cell frequencies were detected in the duodenum and jejunum, respectively. In conclusion, the CG is generally known that it is one of the endocrine markers. Therefore, the abnormality in density of GI endocrine cells detected in the mouse implanted with CT-26 cells may contribute to the development of GI symptoms such as anorexia and indigestion, frequently encountered in patients with cancer.

사자 부신의 형태학적 관찰

박주희,구세광,이재현,이형식,Park, Joo-hee,Ku, Sae-kwang,Lee, Jae-hyun,Lee, Hyeung-sik 대한수의학회 1997 大韓獸醫學會誌 Vol.37 No.3

The morphological and immunohistochemical characteristics of the adrenal gland of the lion(Panthera leo) have been studied. In the light microscopic level, the adrenal gland was surrounded by thick capsules arid weak development of the zona glomerulosa was detected. Some medullary tissues were directly connected with capsules, however the adrenal gland in the lion 'demonstrated somewhat pseudolobulated appearance. Weakly or strongly stained chromogranin-immunoreactive cells were detected in medullary regions of the adrenal gland of the lion by immunohistochemistry, and weakly stained chromogranin-immunoreactive cells were detected more frequent than strongly stained ones.

장기간 위산분비억제제 투여가 혈청 가스트린, 크로모그라닌 A 및 그렐린의 발현에 미치는 영향

김병욱 ( Byung Wook Kim ),이보인 ( Bo In Lee ),김형근 ( Hyung Keun Kim ),조영석 ( Young Seok Cho ),채현석 ( Hiun Suk Chae ),이혜경 ( Hae Kyung Lee ),김현정 ( Hyun Jung Kim ),한석원 ( Sok Won Han ) 대한소화기학회 2009 대한소화기학회지 Vol.53 No.2

목적: 양성자펌프억제제를 장기간 투여하게 되면 위산분비가 억제되어 이차적으로 혈중 가스트린치가 증가하게 된다. 혈중 가스트린치가 증가하면 위의 엔테로크로마핀 유사세포가 과다하게 증식하여 위 유암종 등을 유발할 수 있다. 이번 연구의 목적은 장기간 위산분비억제제를 투여하였을 때, 가스트린, 크로모그라닌 A, 그렐린과 같은 혈중 위단백이 어떻게 변하는지 알아보고자 하였다. 대상 및 방법: 대조군은 6개월 이상 아무런 약물도 투여받지 않는 환자들로 하였고, H2RA군은 6개월 이상 H2 길항제를 복용해오고 있는 환자, PPI군은 6개월 이상 양성자펌프억제제를 복용해오고 있는 환자들을 대상으로 하였다. 대상 환자들에서 공복 시 혈액을 채취하여 RIA법과 ELISA법으로 가스트린, 크로모그라닌 A, 그렐린의 농도를 측정하였다. 결과: 혈청 가스트린과 크로모그라닌 A의 농도가 참고범위 이상을 보인 환자는 대조군과 H2RA군에 비해 PPI군에서 의의있게 높게 발견되었다. 그러나, 혈청 그렐린의 농도는 세 군의 모든 환자에서 참고범위 이내였다. 혈중 가스트린과 크로모그라닌 A농도의 평균치는 대조군과 H2RA군에 비해 PPI군에서 의의있게 증가되어 있었다. 혈중 가스트린과 크로모그라닌 A의 농도 사이에는 의미있는 상관관계를 보였다. 결론: 장기간 H2 길항제 투여는 혈청 가스트린, 크로모그라닌 A의 농도에 영향을 주지 않으나, PPI는 혈청 가스트린과 크로모그라닌 A의 농도를 증가시킨다. 그렐린은 가스트린과 크로모그라닌 A와는 달리 엔테로크로마핀 유사세포를 통하지 않는 다른 방법으로 위산분비에 영향을 주는 것으로 생각한다. Background/Aims: Long-term use of proton pump inhibitor (PPI) induces hypergastrinemia, which results from the suppression of gastric acid secretion. Hypergastrinemia causes enterochromaffin-like (ECL) cell hyperplasia, which is a predisposing factor of carcinoid tumor of stomach. The aim of this study was to identify the effect of long-term gastric acid suppression on the gastric peptides levels, such as gastrin, chromogranin A, or ghrelin. Methods: Control group included patients who had no medication over six months. Both H2RA (H2 receptor antagonist) and PPI groups had medication at least for six months. Fasting blood was taken from each patient to assay serum gastrin, chromogranin A, and ghrelin by RIA and ELISA techniques. Results: The patients with the above reference range of serum gastrin and chromogranin A were more commonly found in PPI group compared to control and H2RA group. However, serum ghrelin level was within the reference range in all the patients regardless of groups. There was no difference in the ratio of serum gastrin/chromogranin A among three groups. Both average serum levels of gastrin and chromogranin A were significantly elevated in PPI group compared to control and H2RA group. There was a significant correlation between the level of serum gastrin and chromogranin A. Conclusions: Long-term administration of H2RA does not affect the serum gastrin and chromogranin A level. However, long-term administration of PPI increases serum gastrin and chromogranin A. Ghrelin may influence gastric acid secretion in other pathway than ECL cell-mediated pathway such as gastrin or chromogranin A. (Korean J Gastroenterol 2009;53:84-89)

생후 1개월령 한국재래산양의 위장관에서 chromogranin A 면역반응세포에 대한 분포 및 출현빈도에 관한 연구

이형식,구세광,이재현 慶山大學校 基礎科學硏究所 2000 基礎科學 Vol.4 No.1

생후 1 개월령의 한국재래산양 위장관에서 chromogranin A 면역반응세포의 각 부위별(위저부, 유문부, 십이지장, 공장, 회장, 결장 및 직장의 7부위) 분포와 출현빈도를 면역조직화학적 방법으로 관찰한 결과, chromogranin A 면역반응세포는 위장관 전부위에서 관찰되었다. 위장관 부위별 출현빈도는 위저부(156.0/1㎟)에서 가장 높은 출현빈도를 나타내었고, 직장(93.0/1㎟), 유문부(84.0/1㎟), 결장(79.0/1㎟), 십이지장(71.0/1㎟), 회장(55.0/1㎟) 및 공장(49.0/1㎟)의 순으로 관찰되었다. 이들 chromogranin A 면역반응세포들은 주로 위저부, 유문부, 십이지장, 회장 및 결장부위에서는 점막샘에 다수, 점막 상피에 소수 관찰되었으며, 공장과 결장에서는 점막 전체에 산재되어 관찰되었다. The regional distribution and relative frequency of chromogranin A-immunoreactive cells in the gastrointestinal tract of the Korean native goat at 1-month-old were studied with immunohistochemical methods. The digestive tract was divided into seven portions, fundus, pylorus, duodenum, jejunum, ileum, colon and rectum. Chromogranin A-immunoreactive cells were detected throughout the whole gastointestinal tract and they were situated to the mucosal glands, which were located in the basal parts of digestive tract, with round, oval and/or spindle shape and surface epithelium with spindle shape. These cells were most predominant in fundus and the relative frequencies of the other portions were observed to be in the following order: rectum > pylorus > colon > duodenum > ileum > jejunum.

생후 6 개월령 한국재래산양의 위장관에서 chromogranin A 면역반응세포에 대한 분포 및 출현빈도에 관한 연구

이형식,이말순 慶山大學校 基礎科學硏究所 2000 基礎科學 Vol.4 No.2

생후 6개월령의 한국재래산양 위장관에서 chromogranin A 면역반응세포의 각 부위별(위저부, 유문부, 십이지장, 공장, 회장, 결장 및 직장의 7부위) 분포와 출현빈도를 면역조직화학적 방법으로 관찰한 결과, chromogranin A 면역반응세포는 위장관 전부위에서 관찰되었다. 위장관 부위별 출현빈도는 위저부(126.0/1㎟)에서 가장 높은 출현빈도를 나타내었고, 유문부(125.0/1㎟), 직장(80.0/1㎟), 결장(70.0/1㎟), 공장(54.0/1㎟), 회장(54.0/1㎟) 및 십이지장(47.0/1㎟)의 순으로 관찰되었다. 이들 chromogranin A 면역반응세포들은 주로 위저부, 유문부, 십이지장, 회장 및 결장부위에서는 점막샘에 다수, 점막 상피에 소수 관찰되었으며, 공장과 결장에서는 점막 전체에 산재되어 관찰되었다. The regional distribution and relative frequency of chromogranin A-immunoreactive cells in the gastrointestinal tract of the Korean native goat at 6-month-old were studied with immunohistochemical methods. The digestive tract was divided into seven portions, fundus, pylorus, duodenum, jejunum, ileum, colon and rectum. Chromogranin A-immunoreactive cells were detected throughout the whole gastrointestinal tract and they were situated to the mucosal glands, which were located in the basal parts of digestive tract, with round, oval and/or spindle shape and surface epithelium with spindle shape. These cells were most predominant in fundus and the relative frequencies of the other portions were observed to be in the following order : fundus > pylorus > rectum > colon >> jejunum = ileum > duodenum.

Circadian variations in salivary chromogranin a concentrations during a 24-hour period in dogs

Kazutaka Kanai,Mariko Hino,Yasutomo Hori,Ruriko Nakao,Fumio Hoshi,Naoyuki Itoh,Seiichi Higuchi 대한수의학회 2008 Journal of Veterinary Science Vol.9 No.4

The purpose of this study was to determine if salivary chromogranin a secretion in dogs exhibits a circadian rhythm. Saliva sampling was performed during three different sessions occurring in three nonconsecutive 24-h periods. Sixteen healthy adult beagle dogs (8 males and 8 females) were moved to a sampling room and housed individually in cages. Saliva samples were obtained every 4 h from 12:00 p.m. to 12:00 p.m. the following day. In the interest of habituation, saliva was obtained hourly from each dog 3 h before the experiment was started. Salivary chromogranin A concentrations were measured using an enzyme-linked immunosorbent assay. No circadian rhythm was detected for salivary chromogranin A secretion, and no differences in salivary chromogranin A concentrations measured every 4 h were demonstrated during the 24-h cycle in dogs.

Subcellular distribution of chromogranins A and B in bovine adrenal chromaffin cells

Huh, Yang Hoon,Bahk, Sook Jin,Ghee, Jung Yeon,Yoo, Seung Hyun Elsevier 2005 FEBS letters Vol.579 No.23

<P><B>Abstract</B></P><P>The major secretory granule proteins chromogranins A (CGA) and B (CGB) have recently been shown to play critical roles in inositol 1,4,5-trisphosphate-dependent intracellular Ca<SUP>2+</SUP> mobilizations. We determined here the subcellular distribution of CGA and CGB based on 3D-images of chromaffin cells, and found that ∼95% of cellular CGA was present in secretory granules while ∼5% was in the endoplasmic reticulum (ER), whereas ∼57% of cellular CGB was in secretory granules while ∼24% and ∼19% were in the ER and nucleus, respectively. These results suggest that chromogranins are at the center of intracellular Ca<SUP>2+</SUP> homeostasis in secretory cells.</P>

한국재래산양 태자의 위장관에서 Chromogranin A 면역반응세포에 대한 분포 및 출현빈도에 관한 연구

이형식,구세광,이재현 慶山大學校 基礎科學硏究所 1997 基礎科學 Vol.1 No.2

임신 180일령의 한국재래산양 태자의 위장관에서 chromogranin A(CgA) 면역반응세포의 각 부위별 출현 분포와 빈도를 면역조직화학적으로 관찰하였던 바, CgA 면역반응세포는 위장관 전 부위에서 관찰되었다. 부위별 분포는 직장(104.0/1㎟)에서 가장 높은 빈도로 출현하였고, 회장(66.6/1㎟), 십이지장(62.1/1㎟), 위저부(53.3/1㎟), 유문부(46.0/1㎟), 결장(34.9/1㎟) 및 공장(30.0/1㎟)의 순으로 관찰되었다. 이들 면역반응세포들은 주로 위저부, 유문부, 십이지장, 회장 및 결장부위에서는 점막샘에 다수, 점막 상피에 소수 분포하였으며, 공장과 결장에서는 점막 전체에 산재되어 있었다. The regional distribution and relative frequency of chromogranin A(CgA)-immunoreactive cells in gastrointestinal tract of the fetus(180 days of gestation) of the Korean native goat were studied with immunohistochemical methods. CgA-immunoreactive cells were detected throughout the gastrointestinal tract and they were situated to the mucosal glands and mucosal epithelia. These cells were most predominant in the rectum(104.0/1㎟), and ileum(66.6/1㎟), duodenum (62.1/1㎟), fundus(53.3/1㎟), pylorus(46.0/1㎟), colon(34.9/1㎟), and jejunum(30.0/1㎟) were followed in that order.