Volume 24 Issue 3
Jun.  2011
Turn off MathJax
Article Contents

Hebron C. CHANG, Daniel R. DOERGE, ChengHong HSIEH, LIN YingJu, FuuJen TSAI. The Covalent Binding of Genistein to the Non-prosthetic-heme-moiety of Bovine Lactoperoxidase Leads to Enzymatic Inactivation[J]. Biomedical and Environmental Sciences, 2011, 24(3): 284-290. doi: 10.3967/0895-3988.2011.03.012
Citation: Hebron C. CHANG, Daniel R. DOERGE, ChengHong HSIEH, LIN YingJu, FuuJen TSAI. The Covalent Binding of Genistein to the Non-prosthetic-heme-moiety of Bovine Lactoperoxidase Leads to Enzymatic Inactivation[J]. Biomedical and Environmental Sciences, 2011, 24(3): 284-290. doi: 10.3967/0895-3988.2011.03.012

The Covalent Binding of Genistein to the Non-prosthetic-heme-moiety of Bovine Lactoperoxidase Leads to Enzymatic Inactivation

doi: 10.3967/0895-3988.2011.03.012
Funds:  the National Science Council,Taiwan(NSC 95-2320-B-408001)%the Interagency Agreement between NCTR/FDA and the National Institute for Environmental Health Sciences/National Toxicology Program,USA
  • Objective Genistein, a major soy isoflavone metabolite (SIF), inactivates oxidation activity of bovine lactoperoxidase (LPO). Modification of the heme moiety of LPO by nitrogen-containing compounds has been shown to inactivate LPO. In contrast, SIF mediated inactivation of LPO does not involve a heme modification and the mechanism of SIF inhibition is poorly understood.Methods After inactivation of LPO by genistein in the presence of H2O2, trypsin-digested LPO peptide fragments were collected and analyzed by MALDI-TOF-MS to characterize the chemical binding of genistein(s) to LPO.Results The heme moiety of LPO was not modified by genistein. A covalent binding study showed that 3H-genistein bound to LPO with a ratio of~12 to 1. After HPLC analysis and peak collection, trypsin-digested peptide fragments were analyzed by MALDI-TOF-MS. The 3H-genistein co-eluted peptide fragments (RT=24 min) were putatively identified as 199IVGYLDEEGVLDQNR214 with two bound genistein molecules or a genistein dimer (2 259 Da), 486TPDNIDIWIGGNAEPMVER504 with two bound genistein molecules or a genistein dimer (2 663 Da), and 161ARWLPAEYEDGLALPFGWTQR182 with three bound genistein molecules or a genistein trimer (3 060 Da). The fragment with a mass of 1 792 Da (RT=36 min) was identified as 132CDENSPYR139 with three genistein molecules or a genistein trimer.Conclusions The results suggest that LPO was inactivated by irreversible covalent binding of genistein or genistein polymers to particular peptide fragments constituting regions of the outward domain. No genistein interaction with the prosthetic heme moiety of LPO was observed.
  • 加载中
  • 加载中
通讯作者: 陈斌, bchen63@163.com
  • 1. 

    沈阳化工大学材料科学与工程学院 沈阳 110142

  1. 本站搜索
  2. 百度学术搜索
  3. 万方数据库搜索
  4. CNKI搜索

Article Metrics

Article views(1133) PDF downloads(17) Cited by()

Proportional views
Related

The Covalent Binding of Genistein to the Non-prosthetic-heme-moiety of Bovine Lactoperoxidase Leads to Enzymatic Inactivation

doi: 10.3967/0895-3988.2011.03.012
Funds:  the National Science Council,Taiwan(NSC 95-2320-B-408001)%the Interagency Agreement between NCTR/FDA and the National Institute for Environmental Health Sciences/National Toxicology Program,USA

Abstract: Objective Genistein, a major soy isoflavone metabolite (SIF), inactivates oxidation activity of bovine lactoperoxidase (LPO). Modification of the heme moiety of LPO by nitrogen-containing compounds has been shown to inactivate LPO. In contrast, SIF mediated inactivation of LPO does not involve a heme modification and the mechanism of SIF inhibition is poorly understood.Methods After inactivation of LPO by genistein in the presence of H2O2, trypsin-digested LPO peptide fragments were collected and analyzed by MALDI-TOF-MS to characterize the chemical binding of genistein(s) to LPO.Results The heme moiety of LPO was not modified by genistein. A covalent binding study showed that 3H-genistein bound to LPO with a ratio of~12 to 1. After HPLC analysis and peak collection, trypsin-digested peptide fragments were analyzed by MALDI-TOF-MS. The 3H-genistein co-eluted peptide fragments (RT=24 min) were putatively identified as 199IVGYLDEEGVLDQNR214 with two bound genistein molecules or a genistein dimer (2 259 Da), 486TPDNIDIWIGGNAEPMVER504 with two bound genistein molecules or a genistein dimer (2 663 Da), and 161ARWLPAEYEDGLALPFGWTQR182 with three bound genistein molecules or a genistein trimer (3 060 Da). The fragment with a mass of 1 792 Da (RT=36 min) was identified as 132CDENSPYR139 with three genistein molecules or a genistein trimer.Conclusions The results suggest that LPO was inactivated by irreversible covalent binding of genistein or genistein polymers to particular peptide fragments constituting regions of the outward domain. No genistein interaction with the prosthetic heme moiety of LPO was observed.

Hebron C. CHANG, Daniel R. DOERGE, ChengHong HSIEH, LIN YingJu, FuuJen TSAI. The Covalent Binding of Genistein to the Non-prosthetic-heme-moiety of Bovine Lactoperoxidase Leads to Enzymatic Inactivation[J]. Biomedical and Environmental Sciences, 2011, 24(3): 284-290. doi: 10.3967/0895-3988.2011.03.012
Citation: Hebron C. CHANG, Daniel R. DOERGE, ChengHong HSIEH, LIN YingJu, FuuJen TSAI. The Covalent Binding of Genistein to the Non-prosthetic-heme-moiety of Bovine Lactoperoxidase Leads to Enzymatic Inactivation[J]. Biomedical and Environmental Sciences, 2011, 24(3): 284-290. doi: 10.3967/0895-3988.2011.03.012

Catalog

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return