Rapid protein identification using N-terminal "sequence tag" and amino acid analysis

Biochemical and Biophysical Research Communications
M R WilkinsA A Gooley

Abstract

Proteins can be identified by amino acid analysis and database matching, but it is often desirable to increase the confidence in identity through the use of other techniques. Here we describe a rapid protein identification method that uses Edman degradation to create a 3 or 4 amino acid N-terminal "sequence tag," following which proteins are subjected to amino acid analysis protein identification procedures. Edman degradation methods have been modified to take only 23 min per cycle, and rapid amino acid analysis techniques are used. The Edman degradation and amino acid analysis is done on a single PVDF membrane-bound protein sample. A computer database matching program is also presented which uses both amino acid composition and "sequence tag" data for protein identification. This method represents the most inexpensive, accurate, and rapid means of protein identification, which is ideal for the screening of proteomes separated by 2-D gel electrophoresis. The creation of N-terminal Edman degradation "sequence-tags" prior to peptide mass fingerprinting of samples should also be useful.

References

Oct 16, 2004·Archives of Oral Biology·Chun-Ming Huang
Dec 1, 1996·Current Biology : CB·M R WilkinsD F Hochstrasser
Jul 31, 2007·Rapid Communications in Mass Spectrometry : RCM·Akihiro ItoShigemi Norioka
Jun 1, 1997·Electrophoresis·A A GooleyK L Williams
Aug 1, 1997·Electrophoresis·I Humphery-SmithW P Blackstock
Sep 18, 1998·Electrophoresis·L TonellaD F Hochstrasser
Mar 1, 1997·Electrophoresis·G L CorthalsA A Gooley
Jun 25, 1998·Electrophoresis·D FenyöB T Chait
Aug 18, 1999·Electrophoresis·M Y HeinkeC G dos Remedios
Jul 5, 2005·The Journal of Biological Chemistry·Frédéric MarinPeter Westbroek
May 1, 2012·Molecular Biotechnology·Daniela QuagliaFrancesca Paradisi
Mar 5, 2002·Microbiology and Molecular Biology Reviews : MMBR·Paul R Graves, Timothy A J Haystead

Related Concepts

Hydrolysis
Sequence Tagged Sites

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.

Sexual Dimorphism in Neurodegeneration

There exist sex differences in neurodevelopmental and neurodegenerative disorders. For instance, multiple sclerosis is more common in women, whereas Parkinson’s disease is more common in men. Here is the latest research on sexual dimorphism in neurodegeneration

HLA Genetic Variation

HLA genetic variation has been found to confer risk for a wide variety of diseases. Identifying these associations and understanding their molecular mechanisms is ongoing and holds promise for the development of therapeutics. Find the latest research on HLA genetic variation here.

Super-resolution Microscopy

Super-resolution microscopy is the term commonly given to fluorescence microscopy techniques with resolutions that are not limited by the diffraction of light. Here are the latest discoveries pertaining to super-resolution microscopy.

Genetic Screens in iPSC-derived Brain Cells

Genetic screening is a critical tool that can be employed to define and understand gene function and interaction. This feed focuses on genetic screens conducted using induced pluripotent stem cell (iPSC)-derived brain cells.

Brain Lower Grade Glioma

Low grade gliomas in the brain form from oligodendrocytes and astrocytes and are the slowest-growing glioma in adults. Discover the latest research on these brain tumors here.

CD4/CD8 Signaling

Cluster of differentiation 4 and 8 (CD8 and CD8) are glycoproteins founds on the surface of immune cells. Here is the latest research on their role in cell signaling pathways.

Alignment-free Sequence Analysis Tools

Alignment-free sequence analyses have been applied to problems ranging from whole-genome phylogeny to the classification of protein families, identification of horizontally transferred genes, and detection of recombined sequences. Here is the latest research.

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.