De Novo Sequence of Novel Protein Based on Chemically-assisted Method Using 4-Sulfophenyl Isothiocyanate Derivatization by Mass Spectrometry

Abstract

Abstract:

Eight tryptic peptides of a novel ginsenoside Rb1 hydrolase(β-glucosidase) purified from Paecilomyces bainier were identified with chemically assisted de novo sequencing followed 4-sulfophenyl isothiocyanate(SPITC) derivatization by MALDI-TOF-TOF mass spectrometry. Some relatively complete sequences were obtained from sulfonated peptides with very low signal/noise ratio. A pair of peptides were identified with non-oxidation and oxidation at methionine, respectively. The results indicated that the sulfonated peptide by SPITC significantly enhanced MALDI-TOF-PSD fragmentation signals and produced a spectrum containing only y⁺ ions. This method facilitated de novo sequencing and spectrum interpretation. By comparing the eight identified peptide sequences with the NCBI database, this purified β-glucosidase proves to be a novel protein that has not yet been reported. Two relatively conserved peptides are remarkable contribution to further research.

Key words: SPITC, MALDI-TOF-TOF MS, Novel protein, De novo sequence

CLC Number:

O657
Q51

TrendMD:

ZHOU Xin-Wen, YAN Qin, ZHOU Pei, YANG Peng-Yuan*. De Novo Sequence of Novel Protein Based on Chemically-assisted Method Using 4-Sulfophenyl Isothiocyanate Derivatization by Mass Spectrometry[J]. Chem. J. Chinese Universities, 2009, 30(4): 706.

[1]	ZHU Ying, LIU Pei, YANG Xiao-Hai, HE Lei-Liang, LI Qing-Zhao, WANG Qing, WANG Ke-Min, HUANG Jin, LIU Jian-Bo. Single-labeled“Turn on” Fluorescent Oligonucleotide Probe for Iodide Detection Based on T-Hg²⁺-T and Inherent Quenching Ability of G-quadruplex [J]. Chem. J. Chinese Universities, 2012, 33(12): 2651.
[2]	SUN Yang, ZHANG Lei, XU Fei, GONG Bo-Lin. Preparation of Chiral Ligand Exchange Chromatographic Stationary Phase by Atom Transfer Radical Polymerization for Separation of Racemates [J]. Chem. J. Chinese Universities, 2012, 33(12): 2644.
[3]	HUANG Jia-Hua, GONG Zhen-Bin, LIN Ji-Jun, DUAN Hua-Ling. Measurement of Complexation Stability Constant and Coordinating Number of Poly(ethylenimine) with Metals [J]. Chem. J. Chinese Universities, 2012, 33(12): 2633.
[4]	SHEN Wei, ZHAO Hong-Mei, LEI Yu-Tao, LING Xiao-Feng, ZHU Qing, ZHANG Ying, ZHANG Yuan-Fu, XU Yi-Zhuang, WU Jin-Guang, WU Yang-Feng, LI Hui-Juan, XU Zhi. Surface Noninvasive Detection of Breast Tumors Using Fourier Transform Infrared Spectroscopy [J]. Chem. J. Chinese Universities, 2012, 33(11): 2390.
[5]	HE Wei-Na, TU Xi-Juan, GUO Xiang-Qun. Spin-Labeled BSA-modified Near-infrared Fluorescent Gold Nanoparticles [J]. Chem. J. Chinese Universities, 2012, 33(11): 2411.
[6]	YANG Jun, QI Li, MA Hui-Min, CHEN Yi. Fabrication of Nylon Membrane-based Microfluidic Chips and Its Application in Color Sensing of Glucose [J]. Chem. J. Chinese Universities, 2012, 33(11): 2405.
[7]	TIAN Qing-Qing, ZENG Wei-Xiu, ZHAO Xin, HOU Yu-Lan, CHEN Bo, YAO Shou-Zhuo. Solvent Induced Phase Separation Extraction-Spectrophotometric Method for Determination of Angiotensin-converting Enzyme Activity [J]. Chem. J. Chinese Universities, 2012, 33(11): 2417.
[8]	FU Hua-Feng, WANG Li-Na, FAN Chun-Ying, LI Xiao-Meng. Rapid Direct Method for Determination of lgP Value by Dispersive Liquid/liquid Microextraction [J]. Chem. J. Chinese Universities, 2012, 33(11): 2435.
[9]	XU Niu-Sheng, YANG Hong-Mei, CUI Meng, SONG Feng-Rui, LIU Zhi-Qiang, LIU Shu-Ying. Investigation of Interaction of Fangchinoline with G-quadruplex DNA by Electrospray Ionization Mass Spectrometry [J]. Chem. J. Chinese Universities, 2012, 33(11): 2430.
[10]	XU Wen, LI Xiao, ZHANG Wei-Ying, YING Xiao-Guang. Preparation and Performance of Sensing Films of Molecularly Imprinted Electrochemical Sensor for L-tryptophan [J]. Chem. J. Chinese Universities, 2012, 33(10): 2199.
[11]	CAO Xia, ZENG Xi, MU Lan, CHEN Yi, WANG Rui-Xiao, ZHANG Yun-Qian, ZHANG Jian-Xin, WEI Gang. Synthesis, Characterization and Aggregation Induced Enhanced Emission of New Phenothiazine Hydrazone [J]. Chem. J. Chinese Universities, 2012, 33(10): 2184.
[12]	WANG Qing, YANG Li-Yan, LIU Jian-Bo, YANG Xiao-Hai, WANG Ke-Min, JI Hai-Ning, LIU Yan. Dynamic Light Scattering for Fluoride Ions Detection [J]. Chem. J. Chinese Universities, 2012, 33(10): 2195.
[13]	HUANG Yan-Qin, QIN Wei-Sheng, REN Hou-Ji, CAO Guo-Yi, LIU Xing-Fen, HUANG Wei. Novel Adenosine Detection Method Based on Cationic Conjugated Polymer and Aptamer [J]. Chem. J. Chinese Universities, 2012, 33(10): 2213.
[14]	EBEYLA Rahman, ASIYA Kerim, PATIME Yasin, PATIMA Nizamidin, ADALAT Abdurahman, ABLIZ Yimit. MB-Stearic Acid Composite Film Optical Waveguide Sensor for the Detection of HCl Gas [J]. Chem. J. Chinese Universities, 2012, 33(10): 2173.
[15]	CUI Zhi-Ping, LIU Shao-Pu, LIU Zhong-Fang, HU Xiao-Li. Dual-wavelength Overlapping Resonance Rayleigh Scattering Spectra of the Interactions of Vitamin B₁₂ with 12-Tungstophosphoric Acid and Their Analytical Applications [J]. Chem. J. Chinese Universities, 2012, 33(10): 2205.

De Novo Sequence of Novel Protein Based on Chemically-assisted Method Using 4-Sulfophenyl Isothiocyanate Derivatization by Mass Spectrometry

PDF (PC)

Knowledge

Cited

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments