2019 ; Vol.29-1: 114~126
|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 Cucurbituril|
J. Microbiol. Biotechnol.2019 ;
|Abstract||This paper introduces three ways to determine host-guest complexation of cucurbituril
(CB) with homocysteine (Hcy). After preincubating Hcy and cysteine (Cys) with CB,
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. Human cystathionine γ-lyase (hCGL) decreased the level of Hcy degradation after
preincubating Hcy and CB. These results suggest that the amount of free Hcy available was
decreased by the formation of a Hcy-CB complex. The immunological signal of anti-Hcy
monoclonal antibody was decreased significantly by preincubating CB 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. Overall, CB can act as a
host by binding selectively with Hcy, but not Cys. The calculated half-complexation formation
concentration of CB 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 host may offer a simple and useful method for determining the Hcy
concentration in plasma or serum.|
|Key_word||Homocysteine, cucurbituril, DTNB, ELISA, human cystathionine γ-lyase|
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