Differentiation of protein species of alpha-2u-globulin according to database entries: A half-theoretical approach

Aksu, Sevil; Tanrikulu, Fatmagül

doi:10.1016/j.jprot.2015.12.024

Cited by 5 publications

(6 citation statements)

References 13 publications

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“…The three proteins were putatively identified as MUP13, Alpha-2u globulin PGCL2 and OBP3. These proteins have also been found in the voided urine of Sprague-Dawley rats, Wistar rats and wild rats [ 21 – 24 , 42 ]. Considering the possible technical difficulties in isolating MUP members and acquiring a single specific MUP [ 26 ], we first investigated OBP3 and MUP13, for which purification has been reported [ 25 , 33 ].…”

Section: Discussionmentioning

confidence: 99%

“…Some studies have efficiently separated MUPs and characterized MUPs related to kinship and individual recognition in mice using IEF [ 10 , 34 ]. Nineteen spots separated by two-dimensional gel electrophoresis (2-DE) were identified as Alpha-2 U - globulin (MUP) using matrix assisted laser desorption ionization-time of flight/time of flight (MALDI-TOF/TOF) mass spectrometry in outbred Wistar rats [ 21 ], and a unique male-specific MUP resolved by narrowed-range IEF was also characterized by MS/ MS in mouse urine [ 44 ]. Inbred Lewis rats may express fewer MUP isoforms than outbred Wistar rats, and thus, IEF is more manageable for separating MUPs from Lewis rats [ 45 ].…”

Section: Discussionmentioning

confidence: 99%

“…Inbred Lewis rats may express fewer MUP isoforms than outbred Wistar rats, and thus, IEF is more manageable for separating MUPs from Lewis rats [ 45 ]. Although more than one MUP is contained in a single band/spot on a 2-DE gel [ 21 ], a greater number of specific unique peptides with high sequence coverage are contributed by the most abundant proteins [ 46 ] and are used for MS-based quantification to successfully identify proteins [ 26 , 47 ]. In our study, the considerable sequence coverage of the matching peptides (82.32%, 78.45% and 75.98% for MUP13, Alpha-2u globulin PGCL2 and OBP3, respectively) was sufficient to demonstrate that these three bands in IEF were the predominant MUPs.…”

Section: Discussionmentioning

confidence: 99%

“…Several MUPs have been characterized from male urine in mice, and some of them have been demonstrated to promote male-male territorial aggression, female attraction, conditioned place preference, or play a role in signaling individual identity [ 4 , 18 – 20 ]. In rats, about 13 MUPs have been found in urine, including OBP3 (PGCL4), MUP13 (PGCL1), and PGCL2 [ 21 – 25 ]. Since the genes of the MUP family share high sequence homology, the purification of MUPs from urine and expression of a single specific protein in vitro is difficult [ 26 ], and the role in intraspecific communication of each MUP isoform in male urine, is rarely experimentally verified in rats [ 25 , 27 ].…”

Section: Introductionmentioning

confidence: 99%

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Two predominant MUPs, OBP3 and MUP13, are male pheromones in rats

Guo

Zhao

et al. 2018

Front Zool

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BackgroundIn rats, urine-borne male pheromones comprise organic volatile compounds and major urinary proteins (MUPs). A number of volatile pheromones have been reported, but no MUP pheromones have been identified in rat urine.ResultsWe used sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing electrophoresis (IEF), nano-liquid chromatography-tandem mass spectrometry (nLC-MS/MS) after in gel digestion of the proteins and quantitative real-time PCR (qRT-PCR) and showed that the levels of two MUPs, odorant-binding protein 3 (OBP3) (i.e. PGCL4) and MUP13 (i.e. PGCL1), in urine and their mRNAs in liver were higher in males than in females and were suppressed by orchidectomy and restored by testosterone treatment (T treatment). We then generated recombinant MUPs (rMUPs) and found that the sexual attractiveness of urine from castrated males to females significantly increased after the addition of either recombinant OBP3 (rOBP3) or recombinant MUP13 (rMUP13). Using c-Fos immunohistochemistry, we further examined neuronal activation in the brains of female rats after they sniffed rOBP3 or rMUP13. Both rOBP3 and rMUP13 activated the accessory olfactory bulb (AOB), medial preoptic area (MPA), bed nucleus of the stria terminalis (BST), medial amygdala (MeA), posteromedial cortical amygdala (PMCo) and ventromedial nucleus of the hypothalamus (VMH), which participate in the neural circuits responsible for pheromone-induced sexual behaviours. In particular, more c-Fos-immunopositive (c-Fos-ir) cells were observed in the posterior AOB than in the anterior AOB.ConclusionsThe expression of OBP3 and MUP13 was male-biased and androgen-dependent. They attracted females and activated brain areas related to sexual behaviours in female rats, suggesting that both OBP3 and MUP13 are male pheromones in rats. Particularly, an OBP excreted into urine was exemplified to be a chemical signal.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Two predominant MUPs, OBP3 and MUP13, are male pheromones in rats

Guo

Zhao

et al. 2018

Front Zool

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“…Although the rat genome sequence was first published in 2004 [27], gene annotation has lagged behind that of the mouse genome and it is more difficult to connect proteins observed in rat urine to the cognate coding sequences predicted from the genome sequence. Furthermore, phenotyping of individual urinary MUPs isoforms in rats has previously been based largely on 1D/2D-SDS-PAGE, or isoelectric focusing (with or without prior purification) [2,3,21,22,[28][29][30][31][32]. Neither PAGE nor isoelectric focusing alone provides adequate resolution for the highly heterogeneous mixture of MUPs isoforms.…”

Section: Introductionmentioning

confidence: 99%

Molecular complexity of the major urinary protein system of the Norway rat,Rattus norvegicus

Gómez‐Baena

Armstrong

Halstead

et al. 2018

Preprint

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26Major urinary proteins (MUP) are the major component of the urinary protein fraction in house mice (Mus 27 spp.) and rats (Rattus spp.). The structure, polymorphism and functions of these lipocalins have been well 28 described in the western European house mouse (Mus musculus domesticus), clarifying their role in 29 semiochemical communication. The complexity of these roles in the mouse raises the question of similar 30 functions in other rodents, including the Norway rat, Rattus norvegicus. Norway rats express MUPs in urine 31 but information about specific MUP isoform sequences and functions is limited. In this study, we present a 32 detailed molecular characterization of the MUP proteoforms expressed in the urine of two laboratory 33 strains, Wistar Han and Brown Norway, and wild caught animals, using a combination of manual gene 34 annotation, intact protein mass spectrometry and bottom-up mass spectrometry-based proteomic 35 approaches. Detailed sequencing of the proteins reveals a less complex pattern of primary sequence 36 polymorphism than the mouse. However, unlike the mouse, rat MUPs exhibit added complexity in the form 37 of post-translational modifications including phosphorylation and exoproteolytic trimming of specific 38 isoforms. The possibility that urinary MUPs may have different roles in rat chemical communication than 39 those they play in the house mouse is also discussed. 40 41 42Physiological production of substantial protein in the urine is well known in both rats and house mice [1, 2]. 43 The protein fraction is dominated by 18-19 kDa, eight stranded beta-barrel lipocalins known as major 44 urinary proteins (MUPs, also named as α2u-globulins when first identified in rats [2, 3]). Urinary MUPs are a 45 heterogeneous mixture of multiple isoforms that are very similar in mass and isoelectric point [4][5][6]. The 46 functions of MUPs have largely been studied in the western European house mouse (Mus musculus 47 domesticus) where they play critical roles in olfactory communication. First, they act as carriers for low 48 molecular weight pheromones and other constituents, delaying their release from urinary scent marks [7][8][9]. 49 MUP polymorphism also provides an identity signal for individual and kin recognition [10][11][12][13][14] and may play a 50 role in species recognition [6]. Finally, MUPs act as pheromones in their own right [14][15][16][17]. In particular, 51 darcin (MUP20, MGI nomenclature; http://www.informatics.jax.org/) has a number of unique properties, 52 including a highly specialized role as a male sex pheromone that also induces competition between males. 53 This protein binds most strongly the abundant volatile male pheromone (S)-2-(sec-butyl)-4,5-54 dihydrothiazole in mouse urine [16][17][18][19]. The pheromonal properties of darcin are retained in the 55 recombinant protein showing that it acts as a pheromone in the absence of bound ligands [16, 17]. 56 57The structure and functions of MUPs in the house mouse are well established and serve to emphasi...

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