A naturally monomeric infrared fluorescent protein for protein labeling in vivo

Dan Yu; Michelle A Baird; John R Allen; Elizabeth S Howe; Matthew P Klassen; Anna Reade; Kalpana Makhijani; Yuanquan Song; Songmei Liu; Zehra Murthy; Shao-Qing Zhang; Orion D Weiner; Thomas B Kornberg; Yuh-Nung Jan; Michael W Davidson; Xiaokun Shu

doi:10.1038/nmeth.3447

A naturally monomeric infrared fluorescent protein for protein labeling in vivo

Nat Methods. 2015 Aug;12(8):763-5. doi: 10.1038/nmeth.3447. Epub 2015 Jun 22.

Authors

Affiliations

¹ 1] Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA. [2] Cardiovascular Research Institute, University of California, San Francisco, San Francisco, California, USA.
² 1] National High Magnetic Field Laboratory, The Florida State University, Tallahassee, Florida, USA. [2] Department of Biological Science, The Florida State University, Tallahassee, Florida, USA.
³ 1] Cardiovascular Research Institute, University of California, San Francisco, San Francisco, California, USA. [2] Department of Physiology, University of California, San Francisco, San Francisco, California, USA. [3] Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, California, USA.
⁴ Cardiovascular Research Institute, University of California, San Francisco, San Francisco, California, USA.
⁵ 1] Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA. [2] Cardiovascular Research Institute, University of California, San Francisco, San Francisco, California, USA. [3] Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania, USA.

Abstract

Infrared fluorescent proteins (IFPs) provide an additional color to GFP and its homologs in protein labeling. Drawing on structural analysis of the dimer interface, we identified a bacteriophytochrome in the sequence database that is monomeric in truncated form and engineered it into a naturally monomeric IFP (mIFP). We demonstrate that mIFP correctly labels proteins in live cells, Drosophila and zebrafish. It should be useful in molecular, cell and developmental biology.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Amino Acid Sequence
Animals
Animals, Genetically Modified
DNA / chemistry
Developmental Biology
Drosophila melanogaster
Fluorescent Dyes / chemistry
Green Fluorescent Proteins / chemistry*
HeLa Cells
Histidine / chemistry
Humans
Infrared Rays*
Luminescent Proteins / chemistry
Mice
Molecular Sequence Data
Mutation
Neurons / metabolism
Plasmids / metabolism
Protein Conformation
Protein Multimerization
Recombinant Fusion Proteins / chemistry
Transfection
Zebrafish

Substances

Fluorescent Dyes
Luminescent Proteins
Recombinant Fusion Proteins
Green Fluorescent Proteins
polyhistidine
Histidine
DNA

Abstract

Publication types

MeSH terms

Substances

Grants and funding