Cloning, expression and purification of the human Islet Amyloid
Polypeptide (hIAPP) from Escherichia coli

J 2015

Cloning, expression and purification of the human Islet Amyloid Polypeptide (hIAPP) from Escherichia coli

CAMARGO, Diana C. Rodrigues, Konstantinos TRIPSIANES, Tobias G. KAPP, Joaquim MENDES, Jasmin SCHUBERT et. al.

Basic information

Original name

Cloning, expression and purification of the human Islet Amyloid Polypeptide (hIAPP) from Escherichia coli

Authors

CAMARGO, Diana C. Rodrigues (276 Germany), Konstantinos TRIPSIANES (300 Greece, guarantor, belonging to the institution), Tobias G. KAPP (276 Germany), Joaquim MENDES (276 Germany), Jasmin SCHUBERT (276 Germany), Burghard CORDES (276 Germany) and Bernd REIF (276 Germany)

Edition

Protein Expression and Purification, SAN DIEGO, Elsevier, 2015, 1046-5928

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

Genetics and molecular biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 1.407

RIV identification code

RIV/00216224:14740/15:00087051

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.1016/j.pep.2014.10.012

UT WoS

000346113600007

Keywords in English

Amyloid; Diabetes; Recombinant protein; human Islet Amyloid Polypeptide (hIAPP); Escherichia coli; Nuclear magnetic resonance (NMR)

Abstract

V originále

Type II diabetes is characterized by deposition of the hormone human Islet Amyloid Polypeptide (hIAPP). Formation of hIAPP amyloid fibrils and aggregates is considered to be responsible for pancreatic beta-cell losses. Therefore, insight into the structure of hIAPP in the solid-state and in solution is of fundamental importance in order to better understand the action of small molecules, which can potentially dissolve protein aggregates and modulate cell toxicity. So far, no procedure has been described that allows to obtain the native human IAPP peptide at high yields. We present here a cloning, expression and purification protocol that permits the production of 2.5 and 3 mg of native peptide per liter of minimal and LB medium, respectively. In the construct, hIAPP is fused to a chitin binding domain (CBD). The CBD is subsequently cleaved off making use of intein splicing reaction which yield amidation of the C-terminus. The N-terminus contains a solubilization domain which is cleaved by V8 protease, avoiding additional residues at the N-terminus. The correct formation of the disulfide bond is achieved by oxidation with H2O2. (C) 2014 Elsevier Inc. All rights reserved.

Citovat

CAMARGO, Diana C. Rodrigues, Konstantinos TRIPSIANES, Tobias G. KAPP, Joaquim MENDES, Jasmin SCHUBERT, Burghard CORDES and Bernd REIF. Cloning, expression and purification of the human Islet Amyloid Polypeptide (hIAPP) from Escherichia coli. Protein Expression and Purification. SAN DIEGO: Elsevier, 2015, vol. 106, Feb, p. 49-56. ISSN 1046-5928. Available from: https://dx.doi.org/10.1016/j.pep.2014.10.012.

@article{1339748,
   author = {Camargo, Diana C. Rodrigues and Tripsianes, Konstantinos and Kapp, Tobias G. and Mendes, Joaquim and Schubert, Jasmin and Cordes, Burghard and Reif, Bernd},
   article_location = {SAN DIEGO},
   article_number = {Feb},
   doi = {http://dx.doi.org/10.1016/j.pep.2014.10.012},
   keywords = {Amyloid; Diabetes; Recombinant protein; human Islet Amyloid Polypeptide (hIAPP); Escherichia coli; Nuclear magnetic resonance (NMR)},
   language = {eng},
   issn = {1046-5928},
   journal = {Protein Expression and Purification},
   title = {Cloning, expression and purification of the human Islet Amyloid Polypeptide (hIAPP) from Escherichia coli},
   url = {http://www.sciencedirect.com/science/article/pii/S1046592814002460},
   volume = {106},
   year = {2015}
}

TY  - JOUR
ID  - 1339748
AU  - Camargo, Diana C. Rodrigues - Tripsianes, Konstantinos - Kapp, Tobias G. - Mendes, Joaquim - Schubert, Jasmin - Cordes, Burghard - Reif, Bernd
PY  - 2015
TI  - Cloning, expression and purification of the human Islet Amyloid Polypeptide (hIAPP) from Escherichia coli
JF  - Protein Expression and Purification
VL  - 106
IS  - Feb
SP  - 49-56
EP  - 49-56
PB  - Elsevier
SN  - 10465928
KW  - Amyloid
KW  - Diabetes
KW  - Recombinant protein
KW  - human Islet Amyloid Polypeptide (hIAPP)
KW  - Escherichia coli
KW  - Nuclear magnetic resonance (NMR)
UR  - http://www.sciencedirect.com/science/article/pii/S1046592814002460
N2  - Type II diabetes is characterized by deposition of the hormone human Islet Amyloid Polypeptide (hIAPP). Formation of hIAPP amyloid fibrils and aggregates is considered to be responsible for pancreatic beta-cell losses. Therefore, insight into the structure of hIAPP in the solid-state and in solution is of fundamental importance in order to better understand the action of small molecules, which can potentially dissolve protein aggregates and modulate cell toxicity. So far, no procedure has been described that allows to obtain the native human IAPP peptide at high yields. We present here a cloning, expression and purification protocol that permits the production of 2.5 and 3 mg of native peptide per liter of minimal and LB medium, respectively. In the construct, hIAPP is fused to a chitin binding domain (CBD). The CBD is subsequently cleaved off making use of intein splicing reaction which yield amidation of the C-terminus. The N-terminus contains a solubilization domain which is cleaved by V8 protease, avoiding additional residues at the N-terminus. The correct formation of the disulfide bond is achieved by oxidation with H2O2. (C) 2014 Elsevier Inc. All rights reserved.
ER  -

CAMARGO, Diana C. Rodrigues, Konstantinos TRIPSIANES, Tobias G. KAPP, Joaquim MENDES, Jasmin SCHUBERT, Burghard CORDES and Bernd REIF. Cloning, expression and purification of the human Islet Amyloid Polypeptide (hIAPP) from Escherichia coli. \textit{Protein Expression and Purification}. SAN DIEGO: Elsevier, 2015, vol.~106, Feb, p.~49-56. ISSN~1046-5928. Available from: https://dx.doi.org/10.1016/j.pep.2014.10.012.

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