The laminin binding protein p40 is involved in inducing limb abnormality of mouse fetuses as the effects of methoxyacetic acid treatment.
This study is intended 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 two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (2-D SDS-PAGE) technique. The 2-D gels reveal one protein with 41.6 kDa and pI 6.4, which expression was downregulated after MAA treatment. Tentative protein identification via peptide mass database search and definitive protein identification via a primary sequence database search indicate 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 mice downregulates the LBP-p40 in the forelimb buds.
Ruyani
Aceng
A
Departemen Biologi, Institut Teknologi Bandung Jl, Ganesha No. 10, Labtek XI, Bandung 40132, Indonesia.
Sudarwati
Sri
S
Sutasurya
Lien A
LA
Sumarsono
Sony H
SH
Gloe
Torsten
T
D000085:Acetates / Q000633:toxicity*; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D004586:Electrophoresis; D004591:Electrophoresis, Polyacrylamide Gel; D005314:Embryonic and Fetal Development / Q000187:drug effects; D005260:Female; D005552:Forelimb / Q000002:abnormalities* / Q000196:embryology / Q000378:metabolism; D007797:Laminin / Q000378:metabolism*; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D011247:Pregnancy
D000085:Acetates; D002352:Carrier Proteins; C488156:LBP-p40 protein, mouse; D007797:Laminin; C013598:methoxyacetic acid