PMID- 29046394
OWN - NLM
STAT- MEDLINE
DCOM- 20171129
LR  - 20171217
IS  - 1470-8728 (Electronic)
IS  - 0264-6021 (Linking)
VI  - 474
IP  - 23
DP  - 2017 Nov 16
TI  - Recombinant human islet amyloid polypeptide forms shorter fibrils and mediates 
      beta-cell apoptosis via generation of oxidative stress.
PG  - 3915-3934
LID - 10.1042/BCJ20170323 [doi]
AB  - Protein misfolding and aggregation play an important role in many human diseases 
      including Alzheimer's, Parkinson's and type 2 diabetes mellitus (T2DM). The human 
      islet amyloid polypeptide (hIAPP) forms amyloid plaques in the pancreas of T2DM 
      subjects (>95%) that are involved in deteriorating islet function and in 
      mediating beta-cell apoptosis. However, the detailed mechanism of action, structure 
      and nature of toxic hIAPP species responsible for this effect remains elusive to 
      date mainly due to the high cost associated with the chemical synthesis of pure 
      peptide required for these studies. In the present work, we attempted to obtain 
      structural and mechanistic insights into the hIAPP aggregation process using 
      recombinant hIAPP (rhIAPP) isolated from Escherichia coli Results from 
      biophysical and structural studies indicate that the rhIAPP self-assembled into 
      highly pure, beta-sheet-rich amyloid fibrils with uniform morphology. 
      rhIAPP-mediated apoptosis in INS-1E cells was associated with increased oxidative 
      stress and changes in mitochondrial membrane potential. The transcript levels of 
      apoptotic genes - Caspase-3 and Bax were found to be up-regulated, while the 
      levels of the anti-apoptotic gene - Bcl2 were down-regulated in rhIAPP-treated 
      cells. Additionally, the expression levels of genes involved in combating 
      oxidative stress namely Catalase, SOD1 and GPx were down-regulated. rhIAPP 
      exposure also affected glucose-stimulated insulin secretion from isolated 
      pancreatic islets. The aggregation of rhIAPP also occurred significantly faster 
      when compared with that of the chemically synthesized peptide. We also show that 
      the rhIAPP fibrils were shorter and more cytotoxic. In summary, our study is one 
      among the few to provide comprehensive evaluation of structural, biophysical and 
      cytotoxic properties of rhIAPP.
CI  - (c) 2017 The Author(s). Published by Portland Press Limited on behalf of the 
      Biochemical Society.
FAU - Dubey, Richa
AU  - Dubey R
AD  - Department of Biosciences and Bioengineering, Indian Institute of Technology, 
      Mumbai 400076, India.
FAU - Minj, Pooja
AU  - Minj P
AD  - Department of Biotechnology, Savitribai Phule Pune University (Formerly 
      University of Pune), Ganeshkhind, Pune, Maharashtra 411007, India.
FAU - Malik, Nikita
AU  - Malik N
AD  - Department of Biosciences and Bioengineering, Indian Institute of Technology, 
      Mumbai 400076, India.
FAU - Sardesai, Devika M
AU  - Sardesai DM
AD  - Department of Biotechnology, Savitribai Phule Pune University (Formerly 
      University of Pune), Ganeshkhind, Pune, Maharashtra 411007, India.
FAU - Kulkarni, Shruti H
AU  - Kulkarni SH
AD  - Department of Biotechnology, Savitribai Phule Pune University (Formerly 
      University of Pune), Ganeshkhind, Pune, Maharashtra 411007, India.
FAU - Acharya, Jhankar D
AU  - Acharya JD
AD  - Department of Zoology, Savitribai Phule Pune University (Formerly University of 
      Pune), Ganeshkhind, Pune, Maharashtra 411007, India.
FAU - Bhavesh, Neel Sarovar
AU  - Bhavesh NS
AD  - Transcription Regulation Group, International Centre for Genetic Engineering and 
      Biotechnology, New Delhi 110067, India.
FAU - Sharma, Shilpy
AU  - Sharma S
AD  - Department of Biotechnology, Savitribai Phule Pune University (Formerly 
      University of Pune), Ganeshkhind, Pune, Maharashtra 411007, India 
      shilpy.sharma@gmail.com ashutoshk@iitb.ac.in.
FAU - Kumar, Ashutosh
AU  - Kumar A
AD  - Department of Biosciences and Bioengineering, Indian Institute of Technology, 
      Mumbai 400076, India shilpy.sharma@gmail.com ashutoshk@iitb.ac.in.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20171116
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (BCL2 protein, human)
RN  - 0 (Islet Amyloid Polypeptide)
RN  - 0 (Proto-Oncogene Proteins c-bcl-2)
RN  - 0 (Recombinant Proteins)
RN  - 0 (SOD1 protein, human)
RN  - 0 (bcl-2-Associated X Protein)
RN  - EC 1.11.1.6 (Catalase)
RN  - EC 1.15.1.1 (Superoxide Dismutase-1)
RN  - EC 3.4.22.- (CASP3 protein, human)
RN  - EC 3.4.22.- (Caspase 3)
SB  - IM
MH  - Apoptosis/*drug effects
MH  - Caspase 3/biosynthesis
MH  - Catalase/biosynthesis
MH  - Cell Line
MH  - Gene Expression Regulation/drug effects
MH  - Humans
MH  - Insulin-Secreting Cells/*metabolism/pathology
MH  - *Islet Amyloid Polypeptide/chemistry/pharmacology
MH  - Oxidative Stress/*drug effects
MH  - Proto-Oncogene Proteins c-bcl-2/biosynthesis
MH  - Recombinant Proteins/chemistry/pharmacology
MH  - Superoxide Dismutase-1/biosynthesis
MH  - bcl-2-Associated X Protein/biosynthesis
OTO - NOTNLM
OT  - amyloid
OT  - apoptosis
OT  - human islet amyloid polypeptide
OT  - solid-state NMR
OT  - type 2 diabetes
EDAT- 2017/10/20 06:00
MHDA- 2017/12/01 06:00
CRDT- 2017/10/20 06:00
PHST- 2017/04/29 00:00 [received]
PHST- 2017/10/09 00:00 [revised]
PHST- 2017/10/11 00:00 [accepted]
PHST- 2017/10/20 06:00 [pubmed]
PHST- 2017/12/01 06:00 [medline]
PHST- 2017/10/20 06:00 [entrez]
AID - BCJ20170323 [pii]
AID - 10.1042/BCJ20170323 [doi]
PST - epublish
SO  - Biochem J. 2017 Nov 16;474(23):3915-3934. doi: 10.1042/BCJ20170323.