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ToxSci Advance Access published online on June 12, 2003
Toxicological Sciences, doi:10.1093/toxsci/kfg159
Toxicological Sciences © Society of Toxicology 2003; all rights reserved
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Received January 21, 2003; accepted May 23, 2003
© 2003 Society of Toxicology

Reproductive and Developmental Toxicology

The Laminin Binding Protein p40 Is Involved in Inducing Limb Abnormality of Mouse Fetuses as the Effects of Methoxyacetic Acid Treatment

Aceng Ruyani 1*, Sri Sudarwati 1, Lien A. Sutasurya 1, Sony H. Sumarsono 1, and Torsten Gloe 2

1 Departemen Biologi, Institut Teknologi Bandung Jl. Ganesha No.10, Labtek XI, Bandung 40132, Indonesia
2 The Institute of Physiology, Ludwig Maximilians University, Schillerstrasse 44, D 80336 Munich, Germany

* To whom correspondence should be addressed. E-mail: Aceng37{at}bi.itb.ac.id.


  Abstract

This study is aimed to characterize a protein that is linked with mouse limb teratogenicity as the effects of methoxyacetic acid (MAA) treatment. A single dose of MAA 10 mmol/kg body weight was given by gavage on gestation day (gd) 11, whereas the control group were administered vehicle only. The pregnant mice were killed at 4 h after MAA treatment, and forelimb buds were isolated from both the control and treated group embryos. Proteins from forelimb buds gd 11+4 h which were precipitated out using 40-60% ammonium sulfate then were analyzed by 2-D SDS-PAGE technique. The 2-D gels reveal one protein with 41.6 kDa and pI 6.4 which expression was down regulated after MAA treatment. Tentative protein identification via peptide mass database search and definitive protein identification via a primary sequence database search indicates that the protein matches exactly to 34/67 kDa laminin binding protein (LBP; P14206, SwissProt), which is encoded by p40 gene (MGI:105381). The identity was further verified by western blotting with an antibody against the 67 kDa LBP. The results suggest that MAA treatment to pregnant mouse down-regulate the LBP-p40 in the forelimb buds.

Key Words: limb teratogenicity, methoxyacetic acid, 2-D SDS-PAGE, LBP-p40 .


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