跳至主要内容

Biochemical and Structural Studies of the Interaction between ARAP1 and CIN85

文献作者:Qingxia Li, Wanfa Yang, Yue Wang, and Wei Liu   《Biochemistry》


ABSTRACTS 

Arf-GAP with Rho-GAP domain, ANK repeat and PH domain-containing protein 1 (ARAP1), Cbl-interacting protein of 85 kDa (CIN85), and casitas B-lineage lymphoma (Cbl) play important roles in epidermal growth factor receptor (EGFR) internalization and recycling. In previous studies, ARAP1 was found to interact with CIN85, and their interaction attenuated the ubiquitination of EGFR. However, the molecular mechanism was still unclear. In this study, we first biochemically and structurally characterized the interaction between ARAP1 and CIN85, and found that the CIN85 SH3B domain bound to the ARAP1 PXPXXRX (except P) XXR/H/K motif with high affinity and specificity. Based on this binding model, we further predicted other potential CIN85 binding partners and tested their interactions biochemically. Moreover, our swapping data and structure alignment analysis suggested that the β2-β3 loops of the CIN85 SH3 domains and the H87ARAP1/E132CIN85 interaction were critical for ARAP1 binding specificity. Finally, our competitive analytical gel-filtration chromatography and isothermal titration calorimetry (ITC) results showed that ARAP1 could compete with Cbl for CIN85 binding, which provides a biochemical basis for the regulatory roles of ARAP1 in the CIN85-mediated EGFR internalizing process.
SCREENSHOT

RELATED PRODUCTS
The ARAP1 aa 80–90 peptide was commercially synthesized by ChinaPeptides Co., Ltd. 
CHAINING
https://pubs.acs.org/doi/abs/10.1021/acs.biochem.8b00057

评论

此博客中的热门博文

Targeting the leptin receptor: To evaluate therapeutic efficacy and anti-tumor effects of Doxil, in vitro and in vivo in mice bearing C26 colon carcinoma tumor

Author:Shahrzad Amiri Darban, Sara Nikoofal-Sahlabadi,  Nafise Amiri, Nafiseh Kiamanesh, Amin Mehrabian, Bamdad Zendehbad, Zahra Gholizadeh, Mahmoud Reza Jaafari     《Colloids and Surfaces B: Biointerfaces》 ABSTRACTS Leptin is an appetite regulatory hormone that is secreted into the blood circulation by the adipose tissue and it functions via its over expressed receptors (Ob-R) in a wide variety of cancers. In the present study, the function of a leptin-derived peptide (LP16, 91–110 of Leptin) was investigated as a targeting ligand to decorate PEGylated liposomal doxorubicin (PLD, Doxil ? ) surface and the anti-tumor activity and therapeutic efficacy of Doxil in C26 (Colon Carcinoma) tumor model were also evaluated. As a result of this, Doxil with different LP16 peptide density (25, 50, 100 and 200 peptide on the surface of each liposome) was successfully prepared and characterized. In vitro results showed significant enhanced cytotoxicity and cellular b...

Deubiquitinating enzyme USP9X regulates cellular clock function by modulating the ubiquitination and degradation of a core circadian protein BMAL1

Author:Yang Zhang, Chunyan Duan, Jing Yang, Suping Chen,  Qing Liu, Liang Zhou, Zhengyun Huang, Ying Xu, Guoqiang Xu     《Biochemical Journal》 ABSTRACTS Living organisms on the earth maintain a roughly 24 h circadian rhythm, which is regulated by circadian clock genes as well as their protein products. Post-translational modifications of core clock proteins could affect the circadian behavior. Although ubiquitination of core clock proteins was studied extensively, the reverse process, deubiquitination, has only begun to unfold and the role of this regulation on circadian function is not completely understood. Here, we use affinity purification and mass spectrometry analysis to identify probable ubiquitin carboxyl-terminal hydrolase FAF-X (USP9X) as an interacting protein of the core clock protein aryl hydrocarbon receptor nuclear translocator-like protein 1 (ARNTL or BMAL1). Through biochemical experiments, we discover that USP9X reduces BMAL1 ubiquitina...

Electrochemical deposition of dopamine–hyaluronic acid conjugates for anti-biofouling bioelectrodes

文献作者:Semin Kim, Yohan Jang, Lindy K. Jang, Sung Hyuk Sunwoo, Tae-il Kim, Seung-Woo Cho and Jae Young Lee     《Journal of Materials Chemistry B》 ABSTRACTS Bioelectrodes have been widely used to effectively mediate electrical signals with biological systems for various biomedical applications, such as biosensors and prosthetic probes. However, the electrical properties of bioelectrodes are frequently degraded in the biological milieu due to biofouling of bioelectrode surfaces. Hence, the development of simple and effective strategies for bioelectrode surface modification is important for the mitigation of biofouling. To this end, we electrochemically modify electrodes with dopamine-conjugated hyaluronic acid (DA–HA); the modified electrodes exhibit highly hydrophilic surfaces. In addition, the electrochemical impedance of the DA–HA-modified electrodes remains similar to those of bare electrodes. The DA–HA-modified electrodes showed reduced non-specific protein ...