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New Insights into the Phylogeny and Gene Context Analysis of Binder of Sperm Proteins (BSPs).

Serrano E, Martínez AB, Arruga D, Pérez-Pé R, Sánchez-Ferrer Á, Muiño-Blanco T, Cebrián-Pérez JA - PLoS ONE (2015)

Bottom Line: Seminal plasma (SP) proteins support the survival of spermatozoa acting not only at the plasma membrane but also by inhibition of capacitation, resulting in higher fertilizing ability.BSPH proteins were found among placental mammals (Eutheria) belonging to the orders Proboscidea, Primates, Lagomorpha, Rodentia, Chiroptera, Perissodactyla and Cetartiodactyla.However, BSPH2 proteins were also found in the Scandentia order and Metatheria clade.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Bioquímica y Biología Molecular y Celular-Instituto Universitario de Investigación en Ciencias Ambientales de Aragón (IUCA), Facultad de Veterinaria, Universidad de Zaragoza, 50013, Zaragoza, Spain.

ABSTRACT
Seminal plasma (SP) proteins support the survival of spermatozoa acting not only at the plasma membrane but also by inhibition of capacitation, resulting in higher fertilizing ability. Among SP proteins, BSP (binder of sperm) proteins are the most studied, since they may be useful for the improvement of semen diluents, storage and subsequent fertilization results. However, an updated and detailed phylogenetic analysis of the BSP protein superfamily has not been carried out with all the sequences described in the main databases. The update view shows for the first time an equally distributed number of sequences between the three families: BSP, and their homologs 1 (BSPH1) and 2 (BSPH2). The BSP family is divided in four subfamilies, BSP1 subfamily being the predominant, followed by subfamilies BSP3, BSP5 and BSP2. BSPH proteins were found among placental mammals (Eutheria) belonging to the orders Proboscidea, Primates, Lagomorpha, Rodentia, Chiroptera, Perissodactyla and Cetartiodactyla. However, BSPH2 proteins were also found in the Scandentia order and Metatheria clade. This phylogenetic analysis, when combined with a gene context analysis, showed a completely new evolutionary scenario for the BSP superfamily of proteins with three defined different gene patterns, one for BSPs, one for BSPH1/BSPH2/ELSPBP1 and another one for BSPH1/BSPH2 without ELSPBP1. In addition, the study has permitted to define concise conserved blocks for each family (BSP, BSPH1 and BSPH2), which could be used for a more reliable assignment for the incoming sequences, for data curation of current databases, and for cloning new BSPs, as the one described in this paper, ram seminal vesicle 20 kDa protein (RSVP20, Ovis aries BSP5b).

No MeSH data available.


In silico analysis of 2FN2 phosphorylcholine binding site.The size of PC binding site 2 is reduced from (A) RSVP20 (BSP5b) to (B) murine BSPH1 and from the latter to (C) murine BSPH2. The first decrease in size is due to a change from a serine (S127 in RSVP20) to a glutamic acid (E109 in mBSPH1). The second decrease in size is due to the change of the latter E109 to a tyrosine (Y104 in mBSPH2). Cartoon representation of the backbone is in tan and surfaces are displayed in CPK (carbon in grey, oxygen in red and nitrogen in blue).
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pone.0137008.g004: In silico analysis of 2FN2 phosphorylcholine binding site.The size of PC binding site 2 is reduced from (A) RSVP20 (BSP5b) to (B) murine BSPH1 and from the latter to (C) murine BSPH2. The first decrease in size is due to a change from a serine (S127 in RSVP20) to a glutamic acid (E109 in mBSPH1). The second decrease in size is due to the change of the latter E109 to a tyrosine (Y104 in mBSPH2). Cartoon representation of the backbone is in tan and surfaces are displayed in CPK (carbon in grey, oxygen in red and nitrogen in blue).

Mentions: Block VI (CxFPFh Y/F, where h stands for hydrophobic amino acid) at the 2FN2 domain is similar to that of Block I (Fig 3A left, cyan). However, this latter block is clearly another fingerprint in BSPH1 proteins (ChFPFWY), where a strictly conserved tryptophan in the middle of β5 appeared, giving rise to a clear hydrophobic tract (V/A FPFW) (Fig 2B). The last hydrophobic amino acid of Block VI (position 52 in Fig 2B, and Y114 in RSVP20; S2 Fig; Fig 3C, cyan) and the last two amino acids of Block VII (GTxxG S/Y/D/E) (positions 64–65 in Fig 2B, G126 and S127 in RSVP20, S2 Fig; Fig 3C, red) form one of the walls of the PC binding pocket 2. The bottom of this pocket 2 is occupied by the completely conserved tryptophan (W71 in Fig 2B, W133 in RSVP20, S2 Fig; Fig 3C, green) of Block VIII (hxxxWCSL T/S). This latter block is also another fingerprint for the BSPH1 family (FGKKWCSLT). Two differences are clearly shown when this block VIII is compared with the homolog block (Block III) in the 1FN2 domain. The first is related with the size, as Block VIII is two amino acids longer than Block III (Fig 2). The second is related with the fact that only one amino acid (W71 in Fig 2B, W133 in RSVP20, S2 Fig) of this Block VIII is involved in the PC binding pocket 2 structure (Fig 3C, green), whereas two amino acids (position 22 and W24 in Fig 2A, R84 and W86 in RSVP20, S2 Fig) of Block III are associated with the PC binding site 1 structure (Fig 3B, green). Finally, Block IX (Y/F N/D xDxxW K/R Y/Q C) at C-ter delimits the opposite wall to Block VI and VII at the PC binding pocket 2 (Fig 3C, magenta). In fact, the wall is formed by the first amino acid of α2 (Y/F N/D x D; position 78 in Fig 2B, F140 in RSVP20, S2 Fig), and the first and the last amino acid of β8 (W K/R Y/Q; W84 and position 86 in Fig 2B, W146 and Y148 in RSVP20, S2 Fig, respectively) (Fig 3C, magenta). This Block IX also differs in size from the homolog in 1FN2, this block again being two amino acids longer (Fig 2). It is noteworthy that this Block IX is an unquestionable BSPH2 fingerprint (YNxDxKWKQC), which has to be supplemented with two strictly conserved amino acids (serine and proline, SP) at the end of the 2FN2 motif (Fig 2B). This conserved C-ter extension after 2FN2 is shown neither in BSPs nor BSPH1s (Fig 2B). The 2FN2 pocket appeared to bind PC with an affinity of -5.29 kcal/mol, which is in the range of murine BSPH1 (-3.5 kcal/mol) but higher than that described for murine BSPH2 (-2.8 kcal/mol) [8]. These docking results could be explained by the decreasing size of the PC binding pocket 2 from BSPs (i.e., RSVP20) to mBSPH1 and from mBSPH1 to mBSPH2 (Fig 4, S2 Fig triangle). The first decrease in size is due to a change from a serine (S127 in RSVP20) to a glutamic acid (E109 in mBSPH1) (Fig 4A and 4B, S2 Fig triangle). The second is due to the change of the latter glutamic acid (E109 in mBSPH1) for a tyrosine (Y104 in mBSPH2), whose electronic density connects with that of Y117, closing the binding site (Fig 4B and 4C, S2 Fig triangle).


New Insights into the Phylogeny and Gene Context Analysis of Binder of Sperm Proteins (BSPs).

Serrano E, Martínez AB, Arruga D, Pérez-Pé R, Sánchez-Ferrer Á, Muiño-Blanco T, Cebrián-Pérez JA - PLoS ONE (2015)

In silico analysis of 2FN2 phosphorylcholine binding site.The size of PC binding site 2 is reduced from (A) RSVP20 (BSP5b) to (B) murine BSPH1 and from the latter to (C) murine BSPH2. The first decrease in size is due to a change from a serine (S127 in RSVP20) to a glutamic acid (E109 in mBSPH1). The second decrease in size is due to the change of the latter E109 to a tyrosine (Y104 in mBSPH2). Cartoon representation of the backbone is in tan and surfaces are displayed in CPK (carbon in grey, oxygen in red and nitrogen in blue).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4557993&req=5

pone.0137008.g004: In silico analysis of 2FN2 phosphorylcholine binding site.The size of PC binding site 2 is reduced from (A) RSVP20 (BSP5b) to (B) murine BSPH1 and from the latter to (C) murine BSPH2. The first decrease in size is due to a change from a serine (S127 in RSVP20) to a glutamic acid (E109 in mBSPH1). The second decrease in size is due to the change of the latter E109 to a tyrosine (Y104 in mBSPH2). Cartoon representation of the backbone is in tan and surfaces are displayed in CPK (carbon in grey, oxygen in red and nitrogen in blue).
Mentions: Block VI (CxFPFh Y/F, where h stands for hydrophobic amino acid) at the 2FN2 domain is similar to that of Block I (Fig 3A left, cyan). However, this latter block is clearly another fingerprint in BSPH1 proteins (ChFPFWY), where a strictly conserved tryptophan in the middle of β5 appeared, giving rise to a clear hydrophobic tract (V/A FPFW) (Fig 2B). The last hydrophobic amino acid of Block VI (position 52 in Fig 2B, and Y114 in RSVP20; S2 Fig; Fig 3C, cyan) and the last two amino acids of Block VII (GTxxG S/Y/D/E) (positions 64–65 in Fig 2B, G126 and S127 in RSVP20, S2 Fig; Fig 3C, red) form one of the walls of the PC binding pocket 2. The bottom of this pocket 2 is occupied by the completely conserved tryptophan (W71 in Fig 2B, W133 in RSVP20, S2 Fig; Fig 3C, green) of Block VIII (hxxxWCSL T/S). This latter block is also another fingerprint for the BSPH1 family (FGKKWCSLT). Two differences are clearly shown when this block VIII is compared with the homolog block (Block III) in the 1FN2 domain. The first is related with the size, as Block VIII is two amino acids longer than Block III (Fig 2). The second is related with the fact that only one amino acid (W71 in Fig 2B, W133 in RSVP20, S2 Fig) of this Block VIII is involved in the PC binding pocket 2 structure (Fig 3C, green), whereas two amino acids (position 22 and W24 in Fig 2A, R84 and W86 in RSVP20, S2 Fig) of Block III are associated with the PC binding site 1 structure (Fig 3B, green). Finally, Block IX (Y/F N/D xDxxW K/R Y/Q C) at C-ter delimits the opposite wall to Block VI and VII at the PC binding pocket 2 (Fig 3C, magenta). In fact, the wall is formed by the first amino acid of α2 (Y/F N/D x D; position 78 in Fig 2B, F140 in RSVP20, S2 Fig), and the first and the last amino acid of β8 (W K/R Y/Q; W84 and position 86 in Fig 2B, W146 and Y148 in RSVP20, S2 Fig, respectively) (Fig 3C, magenta). This Block IX also differs in size from the homolog in 1FN2, this block again being two amino acids longer (Fig 2). It is noteworthy that this Block IX is an unquestionable BSPH2 fingerprint (YNxDxKWKQC), which has to be supplemented with two strictly conserved amino acids (serine and proline, SP) at the end of the 2FN2 motif (Fig 2B). This conserved C-ter extension after 2FN2 is shown neither in BSPs nor BSPH1s (Fig 2B). The 2FN2 pocket appeared to bind PC with an affinity of -5.29 kcal/mol, which is in the range of murine BSPH1 (-3.5 kcal/mol) but higher than that described for murine BSPH2 (-2.8 kcal/mol) [8]. These docking results could be explained by the decreasing size of the PC binding pocket 2 from BSPs (i.e., RSVP20) to mBSPH1 and from mBSPH1 to mBSPH2 (Fig 4, S2 Fig triangle). The first decrease in size is due to a change from a serine (S127 in RSVP20) to a glutamic acid (E109 in mBSPH1) (Fig 4A and 4B, S2 Fig triangle). The second is due to the change of the latter glutamic acid (E109 in mBSPH1) for a tyrosine (Y104 in mBSPH2), whose electronic density connects with that of Y117, closing the binding site (Fig 4B and 4C, S2 Fig triangle).

Bottom Line: Seminal plasma (SP) proteins support the survival of spermatozoa acting not only at the plasma membrane but also by inhibition of capacitation, resulting in higher fertilizing ability.BSPH proteins were found among placental mammals (Eutheria) belonging to the orders Proboscidea, Primates, Lagomorpha, Rodentia, Chiroptera, Perissodactyla and Cetartiodactyla.However, BSPH2 proteins were also found in the Scandentia order and Metatheria clade.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Bioquímica y Biología Molecular y Celular-Instituto Universitario de Investigación en Ciencias Ambientales de Aragón (IUCA), Facultad de Veterinaria, Universidad de Zaragoza, 50013, Zaragoza, Spain.

ABSTRACT
Seminal plasma (SP) proteins support the survival of spermatozoa acting not only at the plasma membrane but also by inhibition of capacitation, resulting in higher fertilizing ability. Among SP proteins, BSP (binder of sperm) proteins are the most studied, since they may be useful for the improvement of semen diluents, storage and subsequent fertilization results. However, an updated and detailed phylogenetic analysis of the BSP protein superfamily has not been carried out with all the sequences described in the main databases. The update view shows for the first time an equally distributed number of sequences between the three families: BSP, and their homologs 1 (BSPH1) and 2 (BSPH2). The BSP family is divided in four subfamilies, BSP1 subfamily being the predominant, followed by subfamilies BSP3, BSP5 and BSP2. BSPH proteins were found among placental mammals (Eutheria) belonging to the orders Proboscidea, Primates, Lagomorpha, Rodentia, Chiroptera, Perissodactyla and Cetartiodactyla. However, BSPH2 proteins were also found in the Scandentia order and Metatheria clade. This phylogenetic analysis, when combined with a gene context analysis, showed a completely new evolutionary scenario for the BSP superfamily of proteins with three defined different gene patterns, one for BSPs, one for BSPH1/BSPH2/ELSPBP1 and another one for BSPH1/BSPH2 without ELSPBP1. In addition, the study has permitted to define concise conserved blocks for each family (BSP, BSPH1 and BSPH2), which could be used for a more reliable assignment for the incoming sequences, for data curation of current databases, and for cloning new BSPs, as the one described in this paper, ram seminal vesicle 20 kDa protein (RSVP20, Ovis aries BSP5b).

No MeSH data available.