PMID- 35417766
OWN - NLM
STAT- MEDLINE
DCOM- 20220509
LR  - 20220617
IS  - 1872-8006 (Electronic)
IS  - 0304-4165 (Linking)
VI  - 1866
IP  - 7
DP  - 2022 Jul
TI  - Developing a novel exenatide-based incretin mimic (alphaB-Ex): Expression, 
      purification and structural-functional characterization.
PG  - 130150
LID - S0304-4165(22)00068-X [pii]
LID - 10.1016/j.bbagen.2022.130150 [doi]
AB  - Among the various treatments, GLP-1 receptor agonists (incretin mimics) such as 
      liraglutide and exenatide have been well received in treating type 2 diabetes 
      mellitus (T2DM) and obesity. In this study, an exenatide analogue, in which 
      methionine at position 14 substituted with leucine, was ligated to human 
      alphaB-crystallin (alphaB-Cry) and then expressed in the bacterial host cells. In the 
      next step, the exenatide analogue was effectively released from the hybrid 
      protein (alphaB-Ex) and subsequently purified using gel filtration chromatography. 
      The HPLC and electrospray ionization mass spectrometry (ESI-MS) analyses 
      respectively suggested a high purity (more than 97%) and an accurate molecular 
      mass for the exenatide analogue (4168.22 Da and 835.01, z = 5). Also, the 
      molecular mass of the alphaB-Ex hybrid protein based on the MALDI-TOF analysis was 
      24,702.162 Da. The secondary structure assessment by the three spectroscopic 
      methods revealed that exenatide analogue and alphaB-Ex hybrid protein have an alpha-helix 
      and a beta-sheet rich structure, respectively. Also, according to the results of the 
      DLS analysis, the alphaB-Ex hybrid protein indicated a high tendency to form large 
      oligomeric structures. The NMR assessment suggested that the hybrid protein 
      exists in its folding state. Both exenatide analogue and the alphaB-Ex hybrid protein 
      revealed a crucial ability to reduce the blood sugar levels in healthy and 
      diabetic mice. They were also capable of inducing insulin secretion to the 
      bloodstream. Overall, our study introduces the alphaB-Ex hybrid protein as a novel 
      incretin mimic, exerting its biological activity for a longer period of time. It 
      might also be considered a potential drug candidate in the treatment of T2DM.
CI  - Copyright (c) 2022. Published by Elsevier B.V.
FAU - Ghanbarnezhad, Mohammad Mehdi
AU  - Ghanbarnezhad MM
AD  - Protein Chemistry Laboratory (PCL), Department of Biology, College of Sciences, 
      Shiraz University, Shiraz, Iran.
FAU - Shahsavani, Mohammad Bagher
AU  - Shahsavani MB
AD  - Protein Chemistry Laboratory (PCL), Department of Biology, College of Sciences, 
      Shiraz University, Shiraz, Iran.
FAU - Mali, Pramod S
AU  - Mali PS
AD  - Department of Biosciences and Bioengineering, IIT Bombay, Powai, Mumbai 400076, 
      India.
FAU - Upadhyay, Mansi
AU  - Upadhyay M
AD  - Department of Biosciences and Bioengineering, IIT Bombay, Powai, Mumbai 400076, 
      India.
FAU - Kumar, Ashutosh
AU  - Kumar A
AD  - Department of Biosciences and Bioengineering, IIT Bombay, Powai, Mumbai 400076, 
      India.
FAU - Albaghlani, Rawayh Muslim
AU  - Albaghlani RM
AD  - Protein Chemistry Laboratory (PCL), Department of Biology, College of Sciences, 
      Shiraz University, Shiraz, Iran.
FAU - Niazi, Ali
AU  - Niazi A
AD  - Institute of Biotechnology, Shiraz University, Shiraz, Iran.
FAU - Yousefi, Reza
AU  - Yousefi R
AD  - Protein Chemistry Laboratory (PCL), Department of Biology, College of Sciences, 
      Shiraz University, Shiraz, Iran; Institute of Biochemistry and Biophysics (IBB), 
      the University of Tehran, Tehran, Iran. Electronic address: 
      ryousefi@shirazu.ac.ir.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20220411
PL  - Netherlands
TA  - Biochim Biophys Acta Gen Subj
JT  - Biochimica et biophysica acta. General subjects
JID - 101731726
RN  - 0 (Incretins)
RN  - 839I73S42A (Liraglutide)
RN  - 9P1872D4OL (Exenatide)
SB  - IM
MH  - Animals
MH  - *Diabetes Mellitus, Experimental
MH  - *Diabetes Mellitus, Type 2/drug therapy
MH  - Exenatide/therapeutic use
MH  - Incretins
MH  - Liraglutide/therapeutic use
MH  - Mice
OTO - NOTNLM
OT  - Exenatide analogue
OT  - Incretin activity
OT  - Mass spectroscopy
OT  - Secondary structure
OT  - Type 2 diabetes mellitus (T2DM)
OT  - alphaB-Ex hybrid protein
EDAT- 2022/04/14 06:00
MHDA- 2022/05/10 06:00
CRDT- 2022/04/13 20:06
PHST- 2022/01/05 00:00 [received]
PHST- 2022/03/30 00:00 [revised]
PHST- 2022/04/07 00:00 [accepted]
PHST- 2022/04/14 06:00 [pubmed]
PHST- 2022/05/10 06:00 [medline]
PHST- 2022/04/13 20:06 [entrez]
AID - S0304-4165(22)00068-X [pii]
AID - 10.1016/j.bbagen.2022.130150 [doi]
PST - ppublish
SO  - Biochim Biophys Acta Gen Subj. 2022 Jul;1866(7):130150. doi: 
      10.1016/j.bbagen.2022.130150. Epub 2022 Apr 11.