Functional Analysis of CXCR3 Splicing Variants and Their Ligands Using NanoBiT-Based Molecular Interaction Assays

황종익,Huong Thi Nguyen,Sunghoon Hurh,Lan Phuong Nguyen,Thai Uy Nguyen,Hee-Kyung Park,Jae Young Seong,Cheol Soon Lee,Byung-Joo Ham 한국분자세포생물학회 2023 Molecules and cells Vol.46 No.5

CXCR3 regulates leukocyte trafficking, maturation, and various pathophysiological conditions. Alternative splicing generates three CXCR3 isoforms in humans. Previous studies investigated the roles of CXCR3 isoforms, and some biochemical data are not correlated with biological relevance analyses. RT-PCR analyses indicate that most cells express all three splicing variants, suggesting that they may mutually affect the chemokine binding and cellular responses of other splicing variants. Here, we performed an integrative analysis of the functional relations among CXCR3 splicing variants and their chemokine-dependent signaling using NanoBiT live cell protein interaction assays. The results indicated that the CXCR3 N-terminal region affected cell surface expression levels and ligand-dependent activation. CXCR3A was efficiently expressed in the plasma membrane and responded to I-TAC, IP-10, and MIG chemokines. By contrast, CXCR3B had low plasma membrane expression and mediated I-TAC–stimulated cellular responses. CXCR3Alt was rarely expressed on the cell surface and did not mediate any cell responses to the tested chemokines; however, CXCR3Alt negatively affected the plasma membrane expression of CXCR3A and CXCR3B and their chemokine-stimulated cellular responses. Jurkat cells express endogenous CXCR3, and exogenous CXCR3A expression enhanced chemotactic activity in response to I-TAC, IP-10, and MIG. By contrast, exogenous expression of CXCR3B and CXCR3Alt eliminated or reduced the CXCR3A-induced chemotactic activity. The PF-4 chemokine did not activate any CXCR3-mediated cellular responses. NanoBiT technology are useful to integrative studies of CXCR3-mediated cell signaling, and expand our knowledge of the cellular responses mediated by molecular interactions among the splicing variants, including cell surface expression, ligand-dependent receptor activation, and chemotaxis.

Inhibitory effects of nafamostat mesilate on proliferation and motility of triple-negative breast cancer cells

황종익,Sunam Mander,박초롱,유동주 한국실험동물학회 2016 한국실험동물학회 학술발표대회 논문집 Vol.2016 No.08

후천진행림프관종 1예(초)

황종익,최윤정,박훈,김진우,유동수 대한피부과학회 2010 초록집 Vol.48 No.10

발목과 손바닥에 발생한 편평황색종 1예(초)

황종익,최윤정,박훈,김진우,유동수 대한피부과학회 2010 초록집 Vol.48 No.10

Apoptotic death of prostate cancer cells by a gonadotropin-releasing hormone-II analog

황종익,박수미,성재영 한국실험동물학회 2013 한국실험동물학회 학술발표대회 논문집 Vol.2013 No.8

급속한 조직확장이 피부편에 미치는 영향에 관한 실험연구

황종익,김태연,정전은 고려대학교 의과대학 1991 고려대 의대 잡지 Vol.28 No.3

Thssue expansion has been a very useful technique to cover the tissue defect with better match in color or texture of the recipient flap and less scar and functional deformity of the donor site .Although this technique has been very popular recently there exist many difficulties with long period of expansion. Many surgeons hesitate rapid expansion in the risk of fear of poor blood flow and harmful histologic change in the expanding skin flap. This study was designed to investigate the effect of the rapid tissue expansion on the skin flap of the rabbits. Rectangualr 200-cc tissue expanders (Dow-Corning Co.) were aseptically placed beneath the skin and panniculus carnosus of 20 New Zealand white rabbits. From the postoperative 2 weeks expansion started with conventional (1-2 week) and rapid (3-4days) interval. In conventional expansion group (Group Ⅰ ) it lasted until 3 months and in rapid expansion group (Group Ⅱ) until 6 weeks. The final inflation volume was 800-1000 cc in both groups. This study compared the rapid expansion versus conventional expansion by the histologic change and quantification of the thickness and the boold flow of the cener in the skin flap above the expander. The results were obtained as followings: 1) Compared with the conventional expansion group, the rapid expansion group reveald slightly inclesed thickness in the epidermis and panniculus carnosus and decreased thickness of the capsule and similiar dermal thickness. 2) No specific difference in histologic change was noted in both groups. 3) Isotope clearance study showed no difference of blood flow between both groups. By the above results, the rapid expansion revealed no harmful effect to the skin flap and further study about the clinical application of the rapid tissue expansion should be done.

신경세포 시냅스에서 Shank2, Homer 1b와 PLC-β3의 신호전달 복합체 형성

황종익,신금주,류성호,서판길 한국뇌학회 2001 한국뇌학회지 Vol.1 No.2

포스포리파아제 C-베타(phospholipase C-β: PLC-β) 동위효소는 G-proteins-coupled receptors(GPCR)와 그의 신호전달 매개체인 heterotrimeric G proteins에 의해 활성화된다. PLC-β는 효소적 활성에 필요한 영역 외에 카르복시 말단에 PSD-95/Dlg/ZO-1 (PDZ)-binding motif를 가진다. 이 motif는 PLC-β가 PDZ domain을 가진 단백질들과 결합함으로써 GPCR 매개 신호전달에서 특이적인 역할을 수행할 가능성을 제시한다. 이에 본 연구자들은 효모 two-hybrid assay를 통하여 Shank2가 PLC-β3와 결합함을 이전에 확인하였다. 본 연구에서는, 면역침전반응을 통해 두 단백질의 결합은 PLC-β3의 카르복시 말단의 PDZ-binding motif를 통하여 이루어짐을 확인하였고 신경세포의 시냅스에서 mGluR1 및 IP3 수용체와 결합하는 Homer 1b 와 Shank2가 PLC-β3와 함께 위치하는 것을 발견하였다. 이러한 결과는 전자현미경을 이용하여 Homer 1b와 Shank2가 위치하는 PSD 영역에서 PLC-β3가 존재함을 확인하고 mGluR1과 Shank2, Homer 1b, PLC-β3가 해마와 소뇌의 많은 부분에서 동시에 발현되는 것을 확인함으로써 다시 한번 입증되었다. 결과를 종합해 볼 때, 이들 세 단백질은 같은 신경세포의 시냅스에서 발현되어 복합체를 형성함으로써 mGluR1에 의해 매개되는 신호전달을 보다 효율적으로 수행할 수 있을 것으로 예상된다. Phospholipase C-β isotypes activated by G protein-coupled receptors (GPCR) and heterotrimeric G proteins carry a PSD-95/Dlg/ZO-1 (PDZ)-binding motif in their carboxyl terminus. This motif may enable PLC-β isotypes to play specific roles in GPCR signaling through interacting with PDZ-containing proteins. We identified Shank2 as a PLC-β3 binding protein, using yeast two-hybrid system in the previous study, and here we characterized the binding of PLC-β3 with Shank2. Immunoprecipitation study showed that Shank2 interacts with PDZ-binding motif of PLC-β3. Moreover, PLC-β3 was colocalized with Shank2 and Homer 1b that interacts with mGluR1 and IP3 receptor on the synapse of primary-cultured neuronal cells. Immunogold EM also showed that PLC-β3 immunoreactivity was detected at the PSD of the CA1 pyramidal neurons. In several regions of hippocampus and cerebellum, the expression pattern of Shank2 was similar to that of mGluR1, Homer 1b, and PLC-β3. These results suggest that PLC-β3, Shank2, and Homer 1b may act as active intermediates in mGluR1-mediated signal transduction by forming signaling complex in neuronal synapses.

정맥 유리 피판술을 이용한 수부 손상 재건술

윤성호,황종익 大韓成形外科學會 1991 Archives of Plastic Surgery Vol.18 No.1

In cases of severe crushing finger injury, skin defect with vascular loss was result in bone, tendon and joint exposures. In patients with certain cases, conventional treatments were vein graft, local flap and disrant flap. Exoposed vein grafred blood vesels were not suitable for graft because of high risk of thrombosis, tissue necrosis and skin graft will not survive on deep structure and local flap may be technically impossible at extensive finger damage. Furthermore distant flap was inappropriate due to postoperative immobilization bulkiness of grafted skin and many times of operations. Therefore in the cases of exposed deep structure, it has been studied about skin flap including subcutaneous vein for a long time. In 1984, Honda et al dad attempte das clinical application and in 1985, Nichtter and Harris et al reported three types of venous flap: 1) total venous perfusion flap 2) arterialized venous perfusion flap 3) total arterial venous perfusion flap. These venous free flaps(skin flaps with subcutaneous veins, flow through venous flap have advantage that can be done without bone shortening as one procedure. And authors did 13 cases of venous flaps, especially 3) and complete survival were 11 cases, partial necrosis were 2 cases.

소아 수지 접합 치험례

김용희,황종익 大韓成形外科學會 1995 Archives of Plastic Surgery Vol.22 No.5

Finger injury in children has been occurred nearly at home and the most frequent cause of injury was door injury and the others were cutter, belt and chain. The level of amputation was Zone I. The treatment methods of children's finger amputation are composite graft, cross finger flap, N-V island flap, V-Y advancement flap, and replantation. But it is difficult for children to maintain the regional flap. And children's fingers are on the way of growing, and injury to the fingers may cause a physical and psychological trauma of children and parents, so we have tried to apply replantation in almost case of amputation in children. Accurate anastomosis, skillful technique, and careful observation were needed for replantation in children because of smaller caliber of vessels. Continuous external bleeding for salvage procedure without excessive bleeding was difficult. Immobilization and careful dressing change were necessary. From March 1989 to December 1992, twenty digits of 17 patients below 15-year-old were replanted. Average age was 5.3-year-old. Frequency of involved digits was 65% in right, 35% in left and was 5% in thumb, 30% in index, 30% in middle, 15% in ring and 20% in tittle finger. After replantation was failed, thenar flap was done in 2 cases. Success of replantation were 12 cases in zone I and 3 cases in zone Ⅲ. In all cases of replantation we treated with hyperbaric oxygen therapy.

수지 수질부 절단상의 재접합술에 대한 임상적 고찰

김용희,이동철,황종익 大韓成形外科學會 1996 Archives of Plastic Surgery Vol.23 No.6

Recent advances in microsurgical techniques and instrument have augmented successful replantation surgery of the amputated body parts. Conventionally, reconstructive methods of the amputated pulp of the digits were used by skin graft, local flap, distant flap (groin flap, cross finger flap, thenar flap etc.) island flap and free flap but, these method would result in complain inadequate color matching, persistant pain, dissensibility, cold intolerance and donor site morbidity. So, we performed replantation when a finger pulp is traumatically amputated or severed at the finger, and we've got superior functional and cosmetic result, than previous modalities of reconstruction of the pulp. From March 1989 to November 1994, 25digits in 23 patient amputated finger pulp were replanted, the venous anastomosis was performed in 12 cases and the others were salvaged via external bleeding technique. The results were as follows ; 88.0% succesibility(22 digit/25 digits), and 3 in 25digit were failed all of those were reconstructed via distant falp and F. T. S. G..