Ultrasensitive detection and characterization of posttranslational modifications using surface-enhanced Raman spectroscopy.
Posttranslational modification (PTM) of proteins is likely to be the most common mechanism of altering the expression of genetic information. It is essential to characterize PTMs to establish a complete understanding of the activities of proteins. Here, we present a sensitive detection method using surface-enhanced Raman spectroscopy (SERS) that can detect PTMs from as little as zeptomoles of peptide. We demonstrate, using model peptides, the ability of SERS to detect a variety of protein modifications, such as acetylation, trimethylation, phosphorylation, and ubiquitination. In addition, we show the capability to obtain positional information for modifications such as trimethylation and phosphorylation using SERS and wavelet decomposition data analysis techniques. We further show that it is possible to apply SERS to detect PTMs from biological samples such as histones. We envision that this detection method might be a valuable technique that is complementary to mass spectrometry in obtaining orthogonal chemical and modification-specific information from biological samples at sensitive levels.
Sundararajan
Narayan
N
Biomedical/Life Sciences, Digital Health Group, Intel Corporation, Santa Clara, California 95052, USA. narayan.sundararajan@intel.com
Mao
Danqian
D
Chan
Selena
S
Koo
Tae-Woong
TW
Su
Xing
X
Sun
Lei
L
Zhang
Jingwu
J
Sung
Kung-bin
KB
Yamakawa
Mineo
M
Gafken
Philip R
PR
Randolph
Tim
T
McLerran
Dale
D
Feng
Ziding
Z
Berlin
Andrew A
AA
Roth
Mark B
MB
D000107:Acetylation; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D006657:Histones / Q000737:chemistry* / Q000378:metabolism*; D008745:Methylation; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D012680:Sensitivity and Specificity; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013499:Surface Properties; D013950:Thymus Gland / Q000737:chemistry / Q000378:metabolism
D006657:Histones