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Molecular heterogeneity of urinary proteins in wild house mouse populations
D H Robertson1, J L Hurst, M S Bolgar
1Department of Biochemistry and Applied Molecular Biology, UMIST, Manchester, UK.
Abstract:
Major urinary proteins (MUPs) from the urine of individual wild mice were characterized using electrospray ionization mass spectrometry (ESI-MS) and compared to MUPs from the urine of inbred mice. The wild mice showed considerable variation between individuals in the expression of a group of MUPs with similar masses. Some individuals excreted MUPs of unique molecular mass whilst some failed to express MUPs seen commonly in the other individuals. All the wild individuals contained proteins not previously observed in inbred mice. Urine from one individual was fractionated using anion exchange chromatography prior to analysis by ESI-MS. By analysing urine from inbred samples under the same conditions it was possible to relate, using mass and net charge in solution, MUPs from the wild sample to the MUPs that have been observed previously in inbred strains. This has allowed tentative identification of some MUPs from the wild mouse. The effect of collection history of urine from wild mice was also investigated. ESI-MS analysis of MUPs in a faecally contaminated sample showed the loss of a C-terminal tripeptide when compared to an uncontaminated sample from the same mouse, consistent with the presence of a specific endopeptidase. Similarly a sample of pooled urine provided by twelve individuals trapped from the same population showed evidence of loss of the C-terminal dipeptide.
Insights
Wild mice exhibit significant individual variation in major urinary proteins (MUPs), with some unique protein expressions and losses observed. These findings reveal novel MUPs and protein modifications in wild mouse populations.
Area of Science:
- Proteomics
- Mammalian Genetics
- Biochemistry
Background:
- Major urinary proteins (MUPs) are a family of rodent pheromone-binding proteins.
- MUPs are highly polymorphic and play roles in social communication.
- Understanding MUPs in wild populations is crucial for evolutionary and ecological studies.
Purpose of the Study:
- To characterize and compare MUPs in wild mice versus inbred laboratory strains.
- To identify novel MUPs and variations in wild mouse populations.
- To investigate the impact of sample collection on MUP integrity.
Main Methods:
- Electrospray ionization mass spectrometry (ESI-MS) for protein characterization.
- Anion exchange chromatography for urine fractionation.
- Comparative analysis of MUPs from wild and inbred mice.
Main Results:
- Wild mice displayed significant inter-individual variation in MUP expression, including unique molecular masses and absence of common MUPs.
- Novel MUPs, not previously observed in inbred mice, were identified in wild individuals.
- Faecal contamination and pooled urine samples indicated C-terminal peptide loss, suggesting enzymatic activity.
Conclusions:
- Wild mouse MUPs exhibit greater diversity than those in inbred strains.
- ESI-MS is effective for identifying MUP variations and post-translational modifications.
- Urine sample integrity is critical for accurate MUP analysis, highlighting potential protease activity in wild populations.
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