Effect of arginine deficiency on arginine-dependent post-translational protein modifications in mice.
Transgenic mice that overexpress arginase-I in their small-intestinal enterocytes suffer from a pronounced, but selective decrease in circulating arginine levels during the suckling period, resulting in impaired growth and development of hair, muscle and immune system. In the present study, we tested the hypothesis that the arginine-deficiency phenotype is caused by arginine-specific post-translational modifications, namely, an increase in the degree of mono-ADP-ribosylation of proteins because of reduced competition by free arginine residues and/or an increase in protein-tyrosine nitration because of an increased O2- production by NO synthases in the presence of limiting amounts of arginine. Arginine ADP-ribosylation and tyrosine nitration of proteins in the affected organs were assayed by Western blot analysis, using specific anti-ADP-ribosylarginine and protein-nitrotyrosine antisera. The composition of the group of proteins that were preferentially arginine ADP-ribosylated or tyrosine-nitrated in the respective organs was strikingly similar. Arginine-deficient mice differed from their controls in a reduced ADP-ribosylation of a 130 kDa and a 65 kDa protein in skin and an increased protein nitration of an 83 kDa protein in bone marrow and a 250 kDa protein in spleen. Since only 20 % of the visualised proteins were differentially modified in a subset of the affected organs, our findings appear to rule out these prominent arginine-dependent post-translational protein modifications as mediators of the characteristic phenotype of severely arginine-deficient mice.
Kwikkers
Karin L
KL
AMC Liver Center and Department of Anatomy and Embryology, Academic Medical Center, University of Amsterdam, Meibergdreef 69-71, 1105 BK, Amsterdam, The Netherlands.
Ruijter
Jan M
JM
Labruyère
Wil T
WT
McMahon
Kathryn K
KK
Lamers
Wouter H
WH
D000244:Adenosine Diphosphate / Q000378:metabolism; D000818:Animals; D001119:Arginase / Q000378:metabolism; D001120:Arginine / Q000172:deficiency* / Q000378:metabolism; D015153:Blotting, Western / Q000379:methods; D001853:Bone Marrow / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D009124:Muscle Proteins / Q000378:metabolism; D010641:Phenotype; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism*; D012867:Skin / Q000378:metabolism; D013154:Spleen / Q000378:metabolism; D013950:Thymus Gland / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism
D009124:Muscle Proteins; D011506:Proteins; D014443:Tyrosine; D000244:Adenosine Diphosphate; D001120:Arginine; D001119:Arginase