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Purification and partial characterization of a new antitumor protein from Tegillarca granosa.

Lv S, Gao J, Liu T, Zhu J, Xu J, Song L, Liang J, Yu R - Mar Drugs (2015)

Bottom Line: Its molecular weight was shown to be 20.320 kDa by ESI-MS/MS, and the isoelectric point of D2-3 was 4.70.The conformational structure of D2-3 was further characterized by UV-vis, FT-IR and CD spectroscopy.Partial amino acid sequences of D2-3 were determined to be LMMTDVEESR, SSHMLSECRRK, KNGRNVDISHKDKG, SSDPTLMDPDDTNKDR, SSDKNTCSKTEYYTR and SSETMPYDVLDTNEMR via MALDI-TOF-MS and de novo sequencing.

View Article: PubMed Central - PubMed

Affiliation: Biotechnological Institute of Chinese Materia Medica, Jinan University, Guangzhou 510632, China. 13691860569@163.com.

ABSTRACT
A new protein, coded as D2-3, was obtained from the marine organism Tegillarca granosa L. by anion exchange and hydrophobic chromatography. The purity of D2-3 was over 99.0% as measured by RP-HPLC. Its molecular weight was shown to be 20.320 kDa by ESI-MS/MS, and the isoelectric point of D2-3 was 4.70. The antitumor activity of D2-3 against four human tumor cell lines was measured by MTT assay. The conformational structure of D2-3 was further characterized by UV-vis, FT-IR and CD spectroscopy. Partial amino acid sequences of D2-3 were determined to be LMMTDVEESR, SSHMLSECRRK, KNGRNVDISHKDKG, SSDPTLMDPDDTNKDR, SSDKNTCSKTEYYTR and SSETMPYDVLDTNEMR via MALDI-TOF-MS and de novo sequencing.

No MeSH data available.


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FT-IR spectrum of D2-3.
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marinedrugs-13-01466-f007: FT-IR spectrum of D2-3.

Mentions: The ultraviolet-visible (UV-vis) absorption, Fourier transform infrared (FT-IR) and circular dichroism (CD) spectroscopies were applied to investigate the structure features of purified protein. The UV-vis absorption spectrum of a 0.1 mg/mL solution of D2-3 in distilled water was determined. D2-3 performed special absorption at 190 nm and 275 nm (Figure 6), which was the typical absorption of peptide bond and amino acid residues such as tyrosine and tryptophan, respectively [18]. FT-IR spectroscopy has been recognized as a valuable and sensitive tool for the examination of protein conformation [19,20]. The amide I band is the sum of all the contributions caused by the secondary of the protein (α-helix, β-sheets, turns and unordered structures) [21]. The FT-IR spectrum of D2-3 is shown in Figure 7. The result indicated that the intense absorption frequencies characterized at 1649.91, 1538.66 and 1307.50 cm−1 were amide I, II and III, respectively. The strong absorption band observed at about 1650 cm−1 was assigned to α-helix [21]. CD spectroscopy provides rapid determinations of protein secondary structure. The CD spectrum (Figure 8) of D2-3 showed two weak negative bands at 208 and 225 nm, and an intense wave at 196 nm. Then the secondary structure of D2-3 was analyzed and calculated using the Jasco protein secondary structure estimation program, and the results showed that it contained 58.2% α-helix, 20.1% β-turn and 21.7% random coil. The total content of α-helix and β-turn accounted for over 70% in the secondary structure. Hence, we regarded D2-3 as a highly ordered and stable protein.


Purification and partial characterization of a new antitumor protein from Tegillarca granosa.

Lv S, Gao J, Liu T, Zhu J, Xu J, Song L, Liang J, Yu R - Mar Drugs (2015)

FT-IR spectrum of D2-3.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4377994&req=5

marinedrugs-13-01466-f007: FT-IR spectrum of D2-3.
Mentions: The ultraviolet-visible (UV-vis) absorption, Fourier transform infrared (FT-IR) and circular dichroism (CD) spectroscopies were applied to investigate the structure features of purified protein. The UV-vis absorption spectrum of a 0.1 mg/mL solution of D2-3 in distilled water was determined. D2-3 performed special absorption at 190 nm and 275 nm (Figure 6), which was the typical absorption of peptide bond and amino acid residues such as tyrosine and tryptophan, respectively [18]. FT-IR spectroscopy has been recognized as a valuable and sensitive tool for the examination of protein conformation [19,20]. The amide I band is the sum of all the contributions caused by the secondary of the protein (α-helix, β-sheets, turns and unordered structures) [21]. The FT-IR spectrum of D2-3 is shown in Figure 7. The result indicated that the intense absorption frequencies characterized at 1649.91, 1538.66 and 1307.50 cm−1 were amide I, II and III, respectively. The strong absorption band observed at about 1650 cm−1 was assigned to α-helix [21]. CD spectroscopy provides rapid determinations of protein secondary structure. The CD spectrum (Figure 8) of D2-3 showed two weak negative bands at 208 and 225 nm, and an intense wave at 196 nm. Then the secondary structure of D2-3 was analyzed and calculated using the Jasco protein secondary structure estimation program, and the results showed that it contained 58.2% α-helix, 20.1% β-turn and 21.7% random coil. The total content of α-helix and β-turn accounted for over 70% in the secondary structure. Hence, we regarded D2-3 as a highly ordered and stable protein.

Bottom Line: Its molecular weight was shown to be 20.320 kDa by ESI-MS/MS, and the isoelectric point of D2-3 was 4.70.The conformational structure of D2-3 was further characterized by UV-vis, FT-IR and CD spectroscopy.Partial amino acid sequences of D2-3 were determined to be LMMTDVEESR, SSHMLSECRRK, KNGRNVDISHKDKG, SSDPTLMDPDDTNKDR, SSDKNTCSKTEYYTR and SSETMPYDVLDTNEMR via MALDI-TOF-MS and de novo sequencing.

View Article: PubMed Central - PubMed

Affiliation: Biotechnological Institute of Chinese Materia Medica, Jinan University, Guangzhou 510632, China. 13691860569@163.com.

ABSTRACT
A new protein, coded as D2-3, was obtained from the marine organism Tegillarca granosa L. by anion exchange and hydrophobic chromatography. The purity of D2-3 was over 99.0% as measured by RP-HPLC. Its molecular weight was shown to be 20.320 kDa by ESI-MS/MS, and the isoelectric point of D2-3 was 4.70. The antitumor activity of D2-3 against four human tumor cell lines was measured by MTT assay. The conformational structure of D2-3 was further characterized by UV-vis, FT-IR and CD spectroscopy. Partial amino acid sequences of D2-3 were determined to be LMMTDVEESR, SSHMLSECRRK, KNGRNVDISHKDKG, SSDPTLMDPDDTNKDR, SSDKNTCSKTEYYTR and SSETMPYDVLDTNEMR via MALDI-TOF-MS and de novo sequencing.

No MeSH data available.


Related in: MedlinePlus