JMB :: Journal of Microbiology and Biotechnology

Author	Se-Ho Park, Jae-Yeul Lee, Hyun-Nam Cho, Kyoung-Ran Kim, Seun-Ah Yang, Hee-Joon Kim, Kwang-Hwan Jhee
Place of duty	Department of Applied Chemistry, Kumoh National Institute of Technology, Gumi 39177, Korea,Institute of Natural Science, Keimyung University, Daegu 42601, Korea
Title	Simple and Novel Assay of the Host-Guest Complexation of Homocysteine with Cucurbit[7]uril
PublicationInfo	J. Microbiol. Biotechnol.2019 ; Vol.29-1
Abstract	This paper introduces three ways to determine host-guest complexation of cucurbit[7]uril (CB[7]) with homocysteine (Hcy). After preincubating Hcy and cysteine (Cys) with CB[7], Ellman’s reagent (DTNB) was used to detect Hcy and Cys. Only Cys reacted with DTNB and Hcy gave a retarded color change. This suggests that the -SH group of Hcy is buried inside CB[7]. Human cystathionine γ-lyase (hCGL) decreased the level of Hcy degradation after preincubating Hcy and CB[7]. These results suggest that the amount of free Hcy available was decreased by the formation of a Hcy-CB[7] complex. The immunological signal of anti-Hcy monoclonal antibody was decreased significantly by preincubating CB[7] with Hcy. The ELISA results also show that ethanethiol group (-CH2CH2SH) of Hcy, which is an epitope of anti-Hcy monoclonal antibody, was blocked by the cavity in CB[7]. Overall, CB[7] can act as a host by binding selectively with Hcy, but not Cys. The calculated half-complexation formation concentration of CB[7] was 58.2 nmol using Ellman’s protocol, 97.9 nmol using hCGL assay and 87.7 nmol using monoclonal antibody. The differing binding abilities of Hcy and Cys towards the CB[7] host may offer a simple and useful method for determining the Hcy concentration in plasma or serum.
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Key_word	Homocysteine, cucurbit[7]uril, DTNB, ELISA, human cystathionine γ-lyase

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