Strain-based Sequence Variations and Structure Analysis of Murine Prostate Specific Spermine Binding Protein Using Mass Spectrometry†

Biochemistry ◽  
2001 ◽  
Vol 40 (32) ◽  
pp. 9725-9733 ◽  
Author(s):  
Pierre Chaurand ◽  
Beverly B. DaGue ◽  
Shuguang Ma ◽  
Susan Kasper ◽  
Richard M. Caprioli
Keyword(s):  
Mass Spectrometry ◽  
Structure Analysis ◽  
Binding Protein ◽  
Sequence Variations

The amino acid sequence of porcine intestinal calcium-binding protein

1979 ◽  
Vol 57 (6) ◽  
pp. 737-748 ◽  
Author(s):  
Theo Hofmann ◽  
Michiko Kawakami ◽  
Anthony J. W. Hitchman ◽  
Joan E. Harrison ◽  
Keith J. Dorrington
Keyword(s):  
Mass Spectrometry ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Intestinal Mucosa ◽  
Calcium Binding ◽  
Binding Protein ◽  
Troponin C ◽  
Calcium Binding Protein ◽  
Mass Spectrometry Analysis ◽  
Spectrometry Analysis

The complete amino acid sequence of the calcium-binding protein (CaBP) from pig intestinal mucosa has been determined: Ac-Ser-Ala-Gln-Lys-Ser-Pro-Ala-Glu-Leu-Lys-Ser-Ile-Phe-Glu-Lys-Tyr-Ala-Ala-Lys-Glu-Gly-Asp-Pro-Asn-Gln-Leu-Ser-Lys-Glu-Glu-Leu-Lys-Gln-Leu-Ile-Gln-Ala-Glu-Phe-Pro-Ser-Leu-Leu-Lys-Gly-Pro-Arg-Thr-Leu-Asp-Asp-Leu-Phe-Gln-Glu-Leu-Asp-Lys-Asn-Gly-Asn-Gly-Glu-Val-Ser-Phe-Glu-Glu-Phe-Gln-Val-Leu-Val-Lys-Lys-Ile-Ser-Gln-OH. The N-terminal octapeptide sequence was determined by mass spectrometry analysis by Morris and Dell. The first 45 residues of bovine CaBP differ only in six positions from the corresponding sequence of the porcine protein, except that the sequence starts in position two of the porcine sequence. The mammalian intestinal CaBP's belong to the troponin-C superfamily on the basis of an analysis by Barker and Dayhoff.

scholarly journals Identification of Vaccinia Virus Replisome and Transcriptome Proteins by Isolation of Proteins on Nascent DNA Coupled with Mass Spectrometry

2017 ◽  
Vol 91 (19) ◽  
Author(s):  
Tatiana G. Senkevich ◽  
George C. Katsafanas ◽  
Andrea Weisberg ◽  
Lisa R. Olano ◽  
Bernard Moss
Keyword(s):  
Mass Spectrometry ◽  
Rna Polymerase ◽  
Vaccinia Virus ◽  
Transcription Initiation ◽  
Nuclear Dna ◽  
Binding Protein ◽  
Viral Proteins ◽  
Animal Cells ◽  
Host Proteins ◽  
Viral Dna

ABSTRACT Poxviruses replicate within the cytoplasm and encode proteins for DNA and mRNA synthesis. To investigate poxvirus replication and transcription from a new perspective, we incorporated 5-ethynyl-2′-deoxyuridine (EdU) into nascent DNA in cells infected with vaccinia virus (VACV). The EdU-labeled DNA was conjugated to fluor- or biotin-azide and visualized by confocal, superresolution, and transmission electron microscopy. Nuclear labeling decreased dramatically after infection, accompanied by intense labeling of cytoplasmic foci. The nascent DNA colocalized with the VACV single-stranded DNA binding protein I3 in multiple puncta throughout the interior of factories, which were surrounded by endoplasmic reticulum. Complexes containing EdU-biotin-labeled DNA cross-linked to proteins were captured on streptavidin beads. After elution and proteolysis, the peptides were analyzed by mass spectrometry to identify proteins associated with nascent DNA. The known viral replication proteins, a telomere binding protein, and a protein kinase were associated with nascent DNA, as were the DNA-dependent RNA polymerase and intermediate- and late-stage transcription initiation and elongation factors, plus the capping and methylating enzymes. These results suggested that the replicating pool of DNA is transcribed and that few if any additional viral proteins directly engaged in replication and transcription remain to be discovered. Among the host proteins identified by mass spectrometry, topoisomerases IIα and IIβ and PCNA were noteworthy. The association of the topoisomerases with nascent DNA was dependent on expression of the viral DNA ligase, in accord with previous proteomic studies. Further investigations are needed to determine possible roles for PCNA and other host proteins detected. IMPORTANCE Poxviruses, unlike many well-characterized animal DNA viruses, replicate entirely within the cytoplasm of animal cells, raising questions regarding the relative roles of viral and host proteins. We adapted newly developed procedures for click chemistry and iPOND (Isolation of proteins on nascent DNA) to investigate vaccinia virus (VACV), the prototype poxvirus. Nuclear DNA synthesis ceased almost immediately following VACV infection, followed swiftly by the synthesis of viral DNA within discrete cytoplasmic foci. All viral proteins known from genetic and proteomic studies to be required for poxvirus DNA replication were identified in the complexes containing nascent DNA. The additional detection of the viral DNA-dependent RNA polymerase and intermediate and late transcription factors provided evidence for a temporal coupling of replication and transcription. Further studies are needed to assess the potential roles of host proteins, including topoisomerases IIα and IIβ and PCNA, which were found associated with nascent DNA.

scholarly journals Structure Analysis of Insoluble Organic Pigments by Mass Spectrometry with Various Ionization Methods

2008 ◽  
Vol 57 (4) ◽  
pp. 265-271 ◽  
Author(s):  
Shigehiro Asai ◽  
Kenichi Fujita ◽  
Masayuki Kubota ◽  
Michiko Kanai ◽  
Hideya Kawasaki ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Structure Analysis ◽  
Organic Pigments ◽  
Ionization Methods

scholarly journals Cross-Linking/Mass Spectrometry Uncovers Details of Insulin-Like Growth Factor Interaction With Insect Insulin Binding Protein Imp-L2

2019 ◽  
Vol 10 ◽  
Author(s):  
Petr Pompach ◽  
Cristina M. Viola ◽  
Jelena Radosavljević ◽  
Jingjing Lin ◽  
Jiří Jiráček ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Growth Factor ◽  
Binding Protein ◽  
Insulin Binding ◽  
Cross Linking ◽  
Insulin Like Growth Factor ◽  
Factor Interaction

scholarly journals Core Structure Analysis of Heavy Oil Components with Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

2019 ◽  
Vol 62 (6) ◽  
pp. 272-281 ◽  
Author(s):  
Shogo TERATANI ◽  
Keita KATANO ◽  
Ryuzo TANAKA ◽  
Tsutomu NAKAMURA ◽  
Hiroshi INOMATA
Keyword(s):  
Mass Spectrometry ◽  
Fourier Transform ◽  
Structure Analysis ◽  
Heavy Oil ◽  
Cyclotron Resonance ◽  
Core Structure ◽  
Ion Cyclotron Resonance ◽  
Ion Cyclotron ◽  
Oil Components

scholarly journals Reaction of Cresyl Saligenin Phosphate, the Organophosphorus Agent Implicated in Aerotoxic Syndrome, with Human Cholinesterases: Mechanistic Studies Employing Kinetics, Mass Spectrometry, and X-ray Structure Analysis

2011 ◽  
Vol 24 (6) ◽  
pp. 797-808 ◽  
Author(s):  
Eugénie Carletti ◽  
Lawrence M. Schopfer ◽  
Jacques-Philippe Colletier ◽  
Marie-Thérèse Froment ◽  
Florian Nachon ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Structure Analysis ◽  
Mechanistic Studies ◽  
X Ray

scholarly journals Structure Analysis and Anti-Tumor and Anti-Angiogenic Activities of Sulfated Galactofucan Extracted from Sargassum thunbergii

Marine Drugs ◽  
2019 ◽  
Vol 17 (1) ◽  
pp. 52 ◽  
Author(s):  
Weihua Jin ◽  
Wanli Wu ◽  
Hong Tang ◽  
Bin Wei ◽  
Hong Wang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Molecular Weight ◽  
High Molecular Weight ◽  
Structure Analysis ◽  
Nmr Spectra ◽  
Low Molecular Weight ◽  
Gel Chromatography ◽  
Sargassum Thunbergii ◽  
Sulfation Pattern ◽  
Fucose Content

Sulfated galactofucan (ST-2) was obtained from Sargassum thunbergii. It was then desulfated to obtain ST-2-DS, and autohydrolyzed and precipitated by ethanol to obtain the supernatant (ST-2-S) and precipitate (ST-2-C). ST-2-C was further fractionated by gel chromatography into two fractions, ST-2-H (high molecular weight) and ST-2-L (low molecular weight). Mass spectrometry (MS) of ST-2-DS was performed to elucidate the backbone of ST-2. It was shown that ST-2-DS contained a backbone of alternating galactopyranose residues (Gal)n (n ≤ 3) and fucopyranose residues (Fuc)n. In addition, ST-2-S was also determined by MS to elucidate the branches of ST-2. It was suggested that sulfated fuco-oligomers might be the branches of ST-2. Compared to the NMR spectra of ST-2-H, the spectra of ST-2-L was more recognizable. It was shown that ST-2-L contain a backbone of (Gal)n and (Fuc)n, sulfated mainly at C4 of Fuc, and interspersed with galactose (the linkages were likely to be 1→2 and 1→6). Therefore, ST-2 might contain a backbone of (Gal)n (n ≤ 3) and (Fuc)n. The sulfation pattern was mainly at C4 of fucopyranose and partially at C4 of galactopyranose, and the branches were mainly sulfated fuco-oligomers. Finally, the anti-tumor and anti-angiogenic activities of ST-2 and its derivates were determined. It was shown that the low molecular-weight sulfated galactofucan, with higher fucose content, had better anti-angiogenic and anti-tumor activities.

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