1. A Strategy for Quality Control of Vespa magnifica (Smith) Venom Based on HPLC Fingerprint Analysis and Multi-Component Separation Combined with Quantitative Analysis
Si-Tong Zhou, Kai Luan, Lian-Li Ni, Ying Wang, Shi-Meng Yuan, Yi-Hao Che, Zi-Zhong Yang, Cheng-Gui Zhang, Zhi-Bin Yang Molecules. 2019 Aug 12;24(16):2920. doi: 10.3390/molecules24162920.
As a folk medicine of the Jingpo minority in Yunnan province, the venom of Vespa magnifica has been commonly used for the treatment of rheumatoid arthritis. Quality standardization of the wasp venom is a necessary step for its pharmaceutical research and development. To control the quality of the wasp venom, a method based on high-performance liquid chromatography (HPLC) was developed for chemical fingerprint analysis. In the chromatographic fingerprinting, chemometrics procedures, including similarity analysis (SA), hierarchical clustering analysis (HCA), and principal component analysis (PCA), were applied to classify 134 batches (S1-S134) of wasp venom from different origins. The HPLC fingerprint method displayed good precision (Relative standard deviation, RSD < 0.27%), stability (in 16 h, RSD < 0.34%), and repeatability (RSD < 1.00%). Simultaneously, four compounds (VMS1, VMS2, VMS3, and VMS4) in the wasp venom were purified and identified. VMS1 was 5-hydroxytryptamine, and the other compounds were three peptides that were sequenced as follows: Gly-Arg-Pro-Hyp-Gly-Phe-Ser-Pro-Phe-Arg-Ile-Asp-NH2 (VMS2), Ile-Asn-Leu-Lys-Ala-Ile-Ala-Ala-Leu-Ala-Lys-Lys-Leu-Leu-NH2 (VMS3), and Phe-Leu-Pro-Ile-Ile-Gly-Lys-Leu-Leu-Ser-Gly-Leu-Leu-NH2 (VMS4). The quantifications for these components were 110.2 mg/g, 26.9 mg/g, 216.3 mg/g, and 58.0 mg/g, respectively. The results of this work indicated that the combination of the chemical fingerprint and quantitative analysis offers a reasonable way to evaluate the quality of wasp venom.
2. The substrate specificity of adenosine 3':5'-cyclic monophosphate-dependent protein kinase of rabbit skeletal muscle
S J Yeaman, P Cohen, D C Watson, G H Dixon Biochem J. 1977 Feb 15;162(2):411-21. doi: 10.1042/bj1620411.
The known amino acid sequences at the two sites on phosphorylase kinase that are phosphorylated by cyclic AMP-dependent protein kinase were extended. The sequences of 42 amino acids around the phosphorylation site on the alpha-subunit and of 14 amino acids around the phosphorylation site on the beta-subunit were shown to be: alpha-subunit Phe-Arg-Arg-Leu-Ser(P)-Ile-Ser-Thr-Glu-Ser-Glx-Pro-Asx-Gly-Gly-His-Ser-Leu-Gly-Ala-Asp-Leu-Met-Ser-Pro-Ser-Phe-Leu-Ser-Pro-Gly-Thr-Ser-Val-Phe(Ser,Pro,Gly)His-Thr-Ser-Lys; beta-subunit, Ala-Arg-Thr-Lys-Arg-Ser-Gly-Ser(P)-VALIle-Tyr-Glu-Pro-Leu-Lys. The sites on histone H2B which are phosphorylated by cyclic AMP-dependent protein kinase in vitro were identified as serine-36 and serine-32. The amino acid sequence in this region is: Lys-Lys-Arg-Lys-Arg-Ser32(P)-Arg-Lys-Glu-Ser36(P)-Tyr-Ser-Val-Tyr-Val- [Iwai, K., Ishikawa, K. & Hayashi, H. (1970) Nature (London) 226, 1056-1058]. Serine-36 was phosphorylated at 50% of the rate at which the beta-subunit of phosphorylase kinase was phosphorylated, and it was phosphorylated 6-7-fold more rapidly than was serine-32. The amino acid sequences when compared with those at the phosphorylation sites of other physiological substrates suggest that the presence of two adjacent basic amino acids on the N-terminal side of the susceptible serine residue may be critical for specific substrate recognition in vivo.
3. A hydrazone-based spectroscopic off-on probe for sensing of basic arginine and lysine
Tianran Wang, Qidan Pang, Zhipu Tong, Hanyue Xiang, Nao Xiao Spectrochim Acta A Mol Biomol Spectrosc. 2021 Sep 5;258:119824. doi: 10.1016/j.saa.2021.119824. Epub 2021 Apr 20.
A simple probe BHN based on naphthol and benzothiazole is reported for detecting of arginine (Arg) and lysine (Lys) with high selectivity and sensitivity. The BHN in aqueous solution upon reacting with Arg or Lys induced a visible color change from colorless to yellow. The probe BHN can also be employed for fluorescence turn-on sensing of Arg and Lys with the limits of detection (LOD) of 5.20 × 10-2 μM and 3.69 × 10-2 μM, respectively. The naked eye colorimetric and fluorimetric detecting is lack of sensitive to other common amino acids including Gly, Ala, Ser, Pro, Val, Thr, Cys, Leu, Ile, Asn, Asp, Glu, Gln, Met, His, and Phe. The sensing mechanism has been proposed by pH investigation and 1H NMR spectra.
