Electrospray ionization mass spectrometry of genetically and chemically modified bacteriorhodopsins

Analytical Biochemistry
P HufnagelD Oesterhelt

Abstract

A simple method for the preparation of bacterio-rhodopsin for mass spectrometry is described, consisting of precipitation of the sample, removal of lipids by washing the sample, and dissolving it in an acidic mixture of organic solvents. Examples demonstrate the method's suitability for the analysis of genetically and chemically modified bacteriorhodopsins. The observed molecular masses are within 0.01% in accordance with the theoretical masses of the wild-type protein and modification products. This allows the detection of modifications accompanied by a mass change greater than 2 Da. Thus, only 9 of 380 possible binary amino acid exchanges are not amenable to analysis. Also chemical modifications of bacteriorhodopsin can be followed since it is possible to characterize reaction products and to approximate yields. However, secondary reactions like hydrolysis in the reaction medium or in the acid solvent used for sample preparation must be considered.

Citations

Jul 9, 1998·Protein Science : a Publication of the Protein Society·J P WhiteleggeK F Faull
Jan 11, 2013·Analytical Chemistry·Julian P Whitelegge
Nov 6, 2002·Proceedings of the National Academy of Sciences of the United States of America·Lorraine B AndersonBridgette A Barry
Sep 15, 1999·Proceedings of the National Academy of Sciences of the United States of America·J P WhiteleggeH R Kaback
Oct 25, 2006·Proceedings of the National Academy of Sciences of the United States of America·Joe CarrollJohn E Walker
Aug 28, 2007·Proceedings of the National Academy of Sciences of the United States of America·Joe CarrollJohn E Walker
May 28, 2011·Trends in Biochemical Sciences·Jeffrey N SavasJohn R Yates
Mar 2, 1999·Journal of Structural Biology·J MayrW Baumeister
Jun 25, 2002·Journal of Molecular Biology·Christopher A PotterMary K Phillips-Jones
Dec 6, 2012·The Journal of Physical Chemistry. B·Annegret P BuschNorbert Hampp
Jun 29, 2006·Molecular & Cellular Proteomics : MCP·Birgit BisleDieter Oesterhelt
Jan 3, 2013·Journal of Mass Spectrometry : JMS·Anil KhanalLars Konermann
Aug 13, 2009·Mass Spectrometry Reviews·Lars KonermannXin Tong
Jan 9, 2010·Rapid Communications in Mass Spectrometry : RCM·Lynsey N JonesAlison E Ashcroft
Nov 25, 2000·Biochimica Et Biophysica Acta·M le MaireJ V Moller
Sep 11, 2002·Journal of Molecular Biology·Thomas FriedrichErnst Bamberg
Jun 5, 2007·Journal of the American Society for Mass Spectrometry·Nina MorgnerBernhard Brutschy
Dec 25, 2004·Proteomics·Christian KleinDieter Oesterhelt
Sep 14, 2004·Journal of Proteome Research·Nan ZhangLiang Li

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.