Transcript Mattow_supplementary_material

SUPPLEMENTARY MATERIAL
Jens Mattow, Ilja Demuth, Gisela Haeselbarth, Peter R. Jungblut,
Joachim Klose
Selenium-binding protein 2,
the major hepatic target for acetaminophen,
shows sex differences in protein abundance
Electrophoresis, submitted.
6805, 6809, 7814*, 8810*
6613*
Parents
4303*
5205*
4301*
5216*
Mother: Calzinato
Father: Aarhus
F1(CxA)
Family 1
5514
Hybrids
Female 1
Female 2
Female 3
Female 4
Female 5
Male 1
Female 6
Mother: Aarhus
Family 2
Parents
Father: Calzinato
F1(AxC)
Females vs. Males
Hybrids
Male 1
Male 2
Male 3
Mother: Calzinato
Father: Aarhus
F1(CxA)
Family 3
Parents
Hybrids
Female 1
Female 2
Female 3
Female 4
Female 5
Female 6
Male 1
Male 2
Mother: Aarhus
Father: Calzinato
F1(AxC)
Hybrids
Female 1
Female 2
Male 1
Male 2
Male 3
Male 4
Male 5
Family 4
Parents
Figure legend
Silver-stained 2-DE patterns of cytosolic liver proteins were investigated in 8
parental mice (4 females; 4 males) of 2 subspecies, namely Mus musculus
musculus (inbred strain: Calcinato B) and Mus musculus domesticus (inbred
strain: Aarhus), as well as in their offspring (25 F1 hybrids; 14 females; 11
males). The primary goal of this analysis was to identify strain-specific protein
variants, which reveal a mode of inheritance compatible with the concept of
genomic imprinting. In this context, the protein patterns of the parental mice
(2 replicate gels per animal) were compared with each other to elucidate strainas well as sex-dependent protein variations.
The figure depicts sectors from the protein patterns of all 33 mice investigated.
The 10 protein spots labeled showed very low intensities in female mice (left
side), but high intensities in males (right side). Seven of these spots (marked by
asterisks) were unambiguously identified as SBP2 by MALDI-MS. The remaining
3 protein spots (SSP5514, SSP6805, SSP6809) most likely represent additional
SBP2 variants, but were not analyzed by MS, due to their absence in preparative
CBB G250 stained 2-DE gels. Edman degradation revealed that SSP6613
represents an N-terminally truncated SBP2 variant, which lacks the amino acids
1-100 of the full-length protein. Similarly, protein spots SSP5216 and SSP5205
were identified as SBP2 variants lacking the amino acids 1-267 and 1-225,
respectively. Edman degradation of SSP8810 did not reveal any sequence data
suggesting that this protein spot represents an N-terminally blocked full-length
SBP2 variant. For reasons of clarity the spot labels are abbreviated, e.g. protein
spot SSP8810 is designated as 8810.
Please note that only the parental mice were analyzed in detail to elucidate sexdependent protein variations. However, reproducible sex differences in SBP2
abundance were also detected in the 25 F1 hybrids.