A study of recombinant human sestrin 1 and sestrin 2 proteins produced in a prokaryotic system

N. Rai; R. Kumar; Md A. Haque; Md I. Hassan; S. Dey

doi:10.1134/S0026893317020170

A study of recombinant human sestrin 1 and sestrin 2 proteins produced in a prokaryotic system

N. Rai, R. Kumar, Md A. Haque, Md I. Hassan, S. Dey

Research output: Contribution to journal › Article › peer-review

Abstract

Sestrins are highly conserved stress-inducible proteins capable of suppressing the production of ROS and signalling through mTORC1. Here we report a study of human sestrin1 (sesn1) and sestrin2 (sesn2) proteins produced in a pET28⁺ vector based prokaryotic system. Mass spectrometry analysis, western blot and surface plasmon resonance (SPR) of affinity purified sesn1 and sesn2 proteins confirmed their identity; biophysical characteristics were observed using circular dichroism (CD) showing that sesn1 and sesn2 have a predominant α-helical structure. Here we describe a simple, one step purification process to purify a large amount of sestrin proteins with significant yield. Further study of recombinant human sestrins may further facilitate the understanding of their roles in eukaryotic cells.

Original language	English (US)
Pages (from-to)	416-425
Number of pages	10
Journal	Molecular Biology
Volume	51
Issue number	3
DOIs	https://doi.org/10.1134/S0026893317020170
State	Published - May 1 2017
Externally published	Yes

Keywords

CD
SPR
prokaryotic system
sestrin

ASJC Scopus subject areas

Biophysics
Structural Biology

Access to Document

10.1134/S0026893317020170

Cite this

@article{83cc19a1af3741098248fc45b3110c95,

title = "A study of recombinant human sestrin 1 and sestrin 2 proteins produced in a prokaryotic system",

abstract = "Sestrins are highly conserved stress-inducible proteins capable of suppressing the production of ROS and signalling through mTORC1. Here we report a study of human sestrin1 (sesn1) and sestrin2 (sesn2) proteins produced in a pET28+ vector based prokaryotic system. Mass spectrometry analysis, western blot and surface plasmon resonance (SPR) of affinity purified sesn1 and sesn2 proteins confirmed their identity; biophysical characteristics were observed using circular dichroism (CD) showing that sesn1 and sesn2 have a predominant α-helical structure. Here we describe a simple, one step purification process to purify a large amount of sestrin proteins with significant yield. Further study of recombinant human sestrins may further facilitate the understanding of their roles in eukaryotic cells.",

keywords = "CD, SPR, prokaryotic system, sestrin",

author = "N. Rai and R. Kumar and Haque, {Md A.} and Hassan, {Md I.} and S. Dey",

note = "Publisher Copyright: {\textcopyright} 2017, Pleiades Publishing, Inc.",

year = "2017",

month = may,

day = "1",

doi = "10.1134/S0026893317020170",

language = "English (US)",

volume = "51",

pages = "416--425",

journal = "Molecular Biology",

issn = "0026-8933",

publisher = "Pleiades Publishing",

number = "3",

}

TY - JOUR

T1 - A study of recombinant human sestrin 1 and sestrin 2 proteins produced in a prokaryotic system

AU - Rai, N.

AU - Kumar, R.

AU - Haque, Md A.

AU - Hassan, Md I.

AU - Dey, S.

PY - 2017/5/1

Y1 - 2017/5/1

N2 - Sestrins are highly conserved stress-inducible proteins capable of suppressing the production of ROS and signalling through mTORC1. Here we report a study of human sestrin1 (sesn1) and sestrin2 (sesn2) proteins produced in a pET28+ vector based prokaryotic system. Mass spectrometry analysis, western blot and surface plasmon resonance (SPR) of affinity purified sesn1 and sesn2 proteins confirmed their identity; biophysical characteristics were observed using circular dichroism (CD) showing that sesn1 and sesn2 have a predominant α-helical structure. Here we describe a simple, one step purification process to purify a large amount of sestrin proteins with significant yield. Further study of recombinant human sestrins may further facilitate the understanding of their roles in eukaryotic cells.

AB - Sestrins are highly conserved stress-inducible proteins capable of suppressing the production of ROS and signalling through mTORC1. Here we report a study of human sestrin1 (sesn1) and sestrin2 (sesn2) proteins produced in a pET28+ vector based prokaryotic system. Mass spectrometry analysis, western blot and surface plasmon resonance (SPR) of affinity purified sesn1 and sesn2 proteins confirmed their identity; biophysical characteristics were observed using circular dichroism (CD) showing that sesn1 and sesn2 have a predominant α-helical structure. Here we describe a simple, one step purification process to purify a large amount of sestrin proteins with significant yield. Further study of recombinant human sestrins may further facilitate the understanding of their roles in eukaryotic cells.

KW - CD

KW - SPR

KW - prokaryotic system

KW - sestrin

UR - http://www.scopus.com/inward/record.url?scp=85021263834&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85021263834&partnerID=8YFLogxK

U2 - 10.1134/S0026893317020170

DO - 10.1134/S0026893317020170

M3 - Article

AN - SCOPUS:85021263834

SN - 0026-8933

VL - 51

SP - 416

EP - 425

JO - Molecular Biology

JF - Molecular Biology

IS - 3

ER -

A study of recombinant human sestrin 1 and sestrin 2 proteins produced in a prokaryotic system

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this