Double quick, double click reversible peptide "stapling".

نویسندگان

  • Claire M Grison
  • George M Burslem
  • Jennifer A Miles
  • Ludwig K A Pilsl
  • David J Yeo
  • Zeynab Imani
  • Stuart L Warriner
  • Michael E Webb
  • Andrew J Wilson
چکیده

The development of constrained peptides for inhibition of protein-protein interactions is an emerging strategy in chemical biology and drug discovery. This manuscript introduces a versatile, rapid and reversible approach to constrain peptides in a bioactive helical conformation using BID and RNase S peptides as models. Dibromomaleimide is used to constrain BID and RNase S peptide sequence variants bearing cysteine (Cys) or homocysteine (hCys) amino acids spaced at i and i + 4 positions by double substitution. The constraint can be readily removed by displacement of the maleimide using excess thiol. This new constraining methodology results in enhanced α-helical conformation (BID and RNase S peptide) as demonstrated by circular dichroism and molecular dynamics simulations, resistance to proteolysis (BID) as demonstrated by trypsin proteolysis experiments and retained or enhanced potency of inhibition for Bcl-2 family protein-protein interactions (BID), or greater capability to restore the hydrolytic activity of the RNAse S protein (RNase S peptide). Finally, use of a dibromomaleimide functionalized with an alkyne permits further divergent functionalization through alkyne-azide cycloaddition chemistry on the constrained peptide with fluorescein, oligoethylene glycol or biotin groups to facilitate biophysical and cellular analyses. Hence this methodology may extend the scope and accessibility of peptide stapling.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Double quick, double click reversible peptide “stapling”† †Electronic supplementary information (ESI) available: Synthesis and characterization, additional biophysical and biochemical analyses. See DOI: 10.1039/c7sc01342f Click here for additional data file. Click here for additional data file. Click here for additional data file.

Synthesis of S peptides......................................................................................................................... 4 WT-S peptide – 8 ................................................................................................................................ 5 OX-S-Cys peptide – 2c ...................................................................................

متن کامل

Investigating peptide sequence variations for 'double-click' stapled p53 peptides.

Stapling peptides for inhibiting the p53/MDM2 interaction is a promising strategy for developing anti-cancer therapeutic leads. We evaluate double-click stapled peptides formed from p53-based diazidopeptides with different staple positions and azido amino acid side-chain lengths, determining the impact of these variations on MDM2 binding and cellular activity. We also demonstrate a K24R mutatio...

متن کامل

Development of a Multifunctional Benzophenone Linker for Peptide Stapling and Photoaffinity Labelling

Photoaffinity labelling is a useful method for studying how proteins interact with ligands and biomolecules, and can help identify and characterise new targets for the development of new therapeutics. We present the design and synthesis of a novel multifunctional benzophenone linker that serves as both a photo-crosslinking motif and a peptide stapling reagent. Using double-click stapling, we at...

متن کامل

Treatment of Colorectal Stricture After Circular Stapling Anastomoses

Performing an end-to-end low rectal anastomosis with the linear single stapling technique was first reported in 1979 (Ravitch & Steichen, 1979). An improved circular stapling technique for anterior resection of the rectum, i.e. double stapling technique, overcoming the problems of insertion of the purse-string on the rectum stump and of disparity in size between the rectum and colon was introdu...

متن کامل

Anastomotic recurrence due to tumor implantation using the double stapling technique.

The double stapling technique is indispensable in low anterior resection for colorectal cancer. However, to prevent local recurrences due to the driving of cancer cells into the anastomosis. Intraluminal lavage should be performed.

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:
  • Chemical science

دوره 8 7  شماره 

صفحات  -

تاریخ انتشار 2017