상세 정보

underline
Metadata Downloads : dc(xml) or Excel
Cited 0 time in scopus ci

Title 

Splitting and self-assembling of far-red fluorescent protein with an engineered beta strand peptide: Application for alpha-synuclein imaging in mammalian cells

Authors 

Joo Oak KeemIn Hwan LeeSun Young KimYongwon JungBong Hyun Chung

Publisher 

Elsevier

Issue Date 

2011

Citation 

Biomaterials, vol. 32, no. 34, pp. 9051-9058

Keywords 

BiosensorsFluorescent proteinMolecular evolutionProtein engineeringSplit protein

Abstract 

We introduce the strategic development of self-assembling peptide/protein fragments based on the far-red fluorescent protein mPlum. The first beta strand (mPlum 1, 18 amino acids) of mPlum was engineered to spontaneously bind with the rest of the protein (mPlum 2-11, next 10 beta strands) and to form a native chromophore. The target beta strand mPlum 1 was separated from mPlum 2-11 and linked via a flexible peptide linker, resulting in fluorescently inactive circularly permuted mPlum protein (CpmPlum). In vitro evolution of this CpmPlum to a fluorescently active form and the subsequent splitting of the engineered mPlum 1 peptide afforded self-assembling mPlum fragments. Recombinantly expressed and synthetically prepared beta strand peptides were successfully assembled with the remaining mPlum protein in vitro and in cells. This developed pair of peptide/protein fragments was effectively used for peptide tag detection of alpha-synuclein in mammalian cells. Sequential expression of self-assembling mPlum fragments offered an entirely genetically encoded sensing system of naturally unfolded alpha-synuclein.

ISSN 

0142-9612

Link 

http://dx.doi.org/10.1016/j.biomaterials.2011.08.029

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


There are no files associated with this item.
qrcode

FusionCharts.
DSpace Software Coptright(c) 2010 MIT and Hewleft-Packard  /  KRIBB-REPOSITORY ( Email:jakim@kribb.re.kr)