Involvement of Complexin 2 in Docking, Locking and Unlocking of Different SNARE Complexes during Sperm Capacitation and Induced Acrosomal Exocytosis
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{"title"=>"Involvement of complexin 2 in docking, locking and unlocking of different SNARE complexes during sperm capacitation and induced acrosomal exocytosis", "type"=>"journal", "authors"=>[{"first_name"=>"Pei Shiue J", "last_name"=>"Tsai", "scopus_author_id"=>"8722306800"}, {"first_name"=>"Ian A.", "last_name"=>"Brewis", "scopus_author_id"=>"7004000450"}, {"first_name"=>"Jillis", "last_name"=>"van Maaren", "scopus_author_id"=>"55062266100"}, {"first_name"=>"Bart M.", "last_name"=>"Gadella", "scopus_author_id"=>"35577972200"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-84857871578", "isbn"=>"1932-6203 (Electronic)\r1932-6203 (Linking)", "doi"=>"10.1371/journal.pone.0032603", "pui"=>"364373490", "sgr"=>"84857871578", "issn"=>"19326203", "pmid"=>"22412896"}, "id"=>"a0bb586b-dd14-322d-a5ab-40b6c40993aa", "abstract"=>"Acrosomal exocytosis (AE) is an intracellular multipoint fusion reaction of the sperm plasma membrane (PM) with the outer acrosomal membrane (OAM). This unique exocytotic event enables the penetration of the sperm through the zona pellucida of the oocyte. We previously observed a stable docking of OAM to the PM brought about by the formation of the trans-SNARE complex (syntaxin 1B, SNAP 23 and VAMP 3). By using electron microscopy, immunochemistry and immunofluorescence techniques in combination with functional studies and proteomic approaches, we here demonstrate that calcium ionophore-induced AE results in the formation of unilamellar hybrid membrane vesicles containing a mixture of components originating from the two fused membranes. These mixed vesicles (MV) do not contain the earlier reported trimeric SNARE complex but instead possess a novel trimeric SNARE complex that contained syntaxin 3, SNAP 23 and VAMP 2, with an additional SNARE interacting protein, complexin 2. Our data indicate that the earlier reported raft and capacitation-dependent docking phenomenon between the PM and OAM allows a specific rearrangement of molecules between the two docked membranes and is involved in (1) recruiting SNAREs and complexin 2 in the newly formed lipid-ordered microdomains, (2) the assembly of a fusion-driving SNARE complex which executes Ca(2+)-dependent AE, (3) the disassembly of the earlier reported docking SNARE complex, (4) the recruitment of secondary zona binding proteins at the zona interacting sperm surface. The possibility to study separate and dynamic interactions between SNARE proteins, complexin and Ca(2+) which are all involved in AE make sperm an ideal model for studying exocytosis.", "link"=>"http://www.mendeley.com/research/involvement-complexin-2-docking-locking-unlocking-different-snare-complexes-during-sperm-capacitatio", "reader_count"=>33, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Professor > Associate Professor"=>3, "Student > Doctoral Student"=>3, "Researcher"=>3, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>1, "Student > Master"=>6, "Other"=>1, "Student > Bachelor"=>2, "Lecturer > Senior Lecturer"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Professor > Associate Professor"=>3, "Student > Doctoral Student"=>3, "Researcher"=>3, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>1, "Student > Master"=>6, "Other"=>1, "Student > Bachelor"=>2, "Lecturer > Senior Lecturer"=>1, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>5, "Agricultural and Biological Sciences"=>18, "Medicine and Dentistry"=>4, "Veterinary Science and Veterinary Medicine"=>2, "Chemistry"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Chemistry"=>{"Chemistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>18}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Unspecified"=>{"Unspecified"=>3}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>2}}, "reader_count_by_country"=>{"Argentina"=>1, "Austria"=>1, "United States"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/671768"], "description"=>"<p>(<b>A</b>) MVs were present around the apical sperm head after AE was induced by calcium ionophore in the absence of bicarbonate. (<b>B</b>) The same was observed for sperm that were first capacitated in presence of bicarbonate and then challenged with Ca<sup>2+</sup> ionophore. (<b>C</b>) MVs isolated from sperm treated as in panel A appeared as unilamellar membrane vesicle structures with a relatively large variation in size (126.7±54.5 nm) and in shape. (<b>D</b>) Unilamellar membrane vesicles isolated from sperm treated as in panel B showed a more homogeneous shape and size (184.4±28.5 nM). These MVs also appear to have more acrosomal matrix components associated to one side of the vesicles (arrow heads) as was already manifest before the MV shedding of the sperm head (see panel B). Statistical analyses on the morphology of MVs present in (C) and (D) were carried out by evaluating three sections from three independent samples (n = 3 values are indicated ± SD). Bar represents 200 nm.</p>", "links"=>[], "tags"=>["morphology", "acrosome", "reacted", "spermatozoa", "subsequently", "formed"], "article_id"=>342245, "categories"=>["Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Pei-Shiue J. Tsai", "Ian A. Brewis", "Jillis van Maaren", "Bart M. Gadella"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0032603.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ultrastructural_morphology_of_acrosome_reacted_spermatozoa_and_the_subsequently_formed_MVs_/342245", "title"=>"Ultrastructural morphology of acrosome reacted spermatozoa and the subsequently formed MVs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 08:05:39"}
  • {"files"=>["https://ndownloader.figshare.com/files/671944"], "description"=>"<p>(<b>A</b>) When MVs were formed in the presence of bicarbonate and Ca<sup>2+</sup> ionophore, a considerable amount of SNARE protein complex was present at 95 kDa (lower panels). A minimal amount of 95 kDa SNARE complexes was observed in MVs from the non-activated sperm when AE was induced by Ca<sup>2+</sup> ionophore in the absence of bicarbonate (upper panels), although a 75 kDa VAMP 2 was found under this bicarbonate depleted condition. Besides the three interacting SNARE proteins found in the MVs, we also identified the capacitation-dependent participation of soluble complexin with the SNARE containing complex. (<b>B</b>) This shows the absence of previously reported SNARE proteins (to some extent syntaxin 1B is present in a 95 kDa complex but syntaxin 2 and VAMP 3 are not participating) in the non-activated MVs indicating that pre-docking and fusion of the acrosome with the PM requires different sets of SNARE proteins. (<b>C</b>) Identification of sperm and testis specific forms of complexin. To demonstrate that the unexpected 40 kDa complexin was not an artifact, whole sperm lysates and a mouse testis tissue homogenate were used. For the comparisons between different tissues, both pig and mouse brain homogenate were included. Despite the presence of multiple complexin positive bands, we found in both mouse and pig brain, a huge amount of complexin monomer was present. In contrast to the specific detection of 40 and 79 kDa complexin (marked with asterisks) in the sperm and testis, no 40 kDa complexin can be detected in the brain tissue of both species suggesting this unexpected 40 kDa complexin is a sperm-specific form rather than the artifact. A total of 35 µg crude protein was loaded to each lane of these Western blots from: 1: whole sperm cell lysate; 2: the remaining sperm head fraction (in the absence of MVs); 3: MVs. Samples were either incubated at the RT in the absence of reducing agent (non-reducing conditions) or boiled at 100°C for 10 min in the presence of 0.1 M DTT (reducing conditions). The arrows indicate the expected molecular weight of protein on the Western blot.</p>", "links"=>[], "tags"=>["ionophore", "incubation", "mvs", "amounts", "snare"], "article_id"=>342429, "categories"=>["Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Pei-Shiue J. Tsai", "Ian A. Brewis", "Jillis van Maaren", "Bart M. Gadella"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0032603.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Bicarbonate_and_Ca_2_ionophore_incubation_results_in_MVs_that_contain_high_amounts_of_SNARE_protein_complexes_/342429", "title"=>"Bicarbonate and Ca<sup>2+</sup> ionophore incubation results in MVs that contain high amounts of SNARE protein complexes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 08:06:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/672074"], "description"=>"<p>(<b>A–D</b>) Immunofluorescence detection of SNARE interacting proteins complexin 1/2, synaptotagmin 4 and Munc 18-2 showed a homogenous distribution over the entire sperm head in the control, non-activated sperm (B1–D1). Munc 18-2 had an additional strong staining at the post-equatorial region (D1, indicated with arrow heads). Upon capacitation, these proteins relocalized to the apical sperm head (marked with asterisk) and appeared as punctate aggregates indicating their association with other proteins (B2–D2). When AE was induced (either in the absence [B3–D3] or in the presence [B4–D4] of bicarbonate), the majority of the signal at the apical ridge of the sperm head disappeared, which was due to the removal of apical membranes after shedding MVs from the sperm heads. A minor signal can still be detected in some sperm cells when AE is induced in the absence of bicarbonate while the majority of the signal was lost in the acrosome reacted sperm in the presence of bicarbonate; this is due to different efficiencies in AE induction. Purified rabbit IgG were used as a negative control (A1–A4) since these Abs were raised in rabbit. To demonstrate the specificity of the signals observed, sperm cells were also incubated in the absence of primary Ab (A5) or were incubated in the primary Ab that was immunized with specific blocking peptide of which the antibody was produced (B5–D5). Bars represent 10 µm. (<b>E</b>) Validation the presence of Munc18-2 and synaptotagmin 4 in porcine sperm. Whole sperm cell lysates (WC) from 4 different incubation conditions (G1–G4, see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032603#s4\" target=\"_blank\">Methods and Materials</a>) were used for the further validation on the presence of Munc 18-2 and synaptotagmin 4 observed in other independent experiments. The arrows indicate the expected molecular weight of protein on the Western blot.</p>", "links"=>[], "tags"=>["relocalization", "snare", "interacting"], "article_id"=>342569, "categories"=>["Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Pei-Shiue J. Tsai", "Ian A. Brewis", "Jillis van Maaren", "Bart M. Gadella"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0032603.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Bicarbonate_dependent_relocalization_of_SNARE_interacting_proteins_/342569", "title"=>"Bicarbonate-dependent relocalization of SNARE interacting proteins.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 08:07:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/672245"], "description"=>"<p>(<b>A</b>) Dot blots and density graphs for flotillin 1 in detergent treated sperm samples after separation over a discontinuous sucrose gradient. Detergent resistant membranes (DRM) were isolated using Triton X-100. DRMs were located at fraction 4–6 based on the enrichment of the raft marker protein flotillin 1. (<b>B</b>) Western blots of the gradient fractions 1–13 for complexin 1/2 under reducing conditions. Two monomeric forms (indicated with the arrow head) of complexin were detected at 19 (complexin 1) and 20 kDa (complexin 2) in porcine sperm. Both complexins were predominately present in the soluble fraction, no migration or complexed complexin can be detected in the control non-activated sperm (i.e. in the absence of bicarbonate; upper panel). When sperm cells were capacitated in the presence of bicarbonate, a clear migration of complexin 2 from the non-DRM fractions into the DRM fractions was observed suggesting the capacitation-dependent aggregation of complexin 2 to the apical area of the sperm head (middle panel) which remains in the DRM after the acrosome reaction (lower panel). (<b>C</b>) See (B) but these Western blots were performed on sample separated by SDS-PAGE under non-reducing conditions. Complexin 2 remains associated with different protein (complexes) than only with the trimeric SNARE complex. The 40 kDa complexin positive signal (indicated with an asterisk) indicated a specific dimer form of complexin in the porcine sperm. Further identification of 79 kDa complexin (marked with double asterisks) may reflect to the complexin-SNAREpin complex that does not have the full fusion ability and may mirror the complexin sub-population observed at the post-equatorial region of the sperm head in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032603#pone-0032603-g003\" target=\"_blank\">Figure 3</a>. A major shift of the signal from 79 kDa to 95 kDa upon AE indicated the participation of the R-SNARE protein in these complexes.</p>", "links"=>[], "tags"=>["induces", "complexin", "snare", "raft", "non-raft"], "article_id"=>342729, "categories"=>["Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Pei-Shiue J. Tsai", "Ian A. Brewis", "Jillis van Maaren", "Bart M. Gadella"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0032603.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Capacitation_induces_complexin_2_interaction_with_the_SNARE_complex_in_both_the_raft_and_non_raft_fractions_/342729", "title"=>"Capacitation induces complexin 2 interaction with the SNARE complex in both the raft and non-raft fractions.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 08:08:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/672358"], "description"=>"<p>(<b>A</b>) A Q-SNARE complex containing syntaxin 3 and SNAP 23 was detected on Western blots of the high sucrose gradient fractions (#11–13, indicated with number signs). This complex was found to lack the complementary R-SNARE VAMP 2 and thus enabled us to distinguish two different SNARE containing complexes in the porcine sperm. (<b>B</b>) The previously identified SNARE complex responsible for acrosome to plasma membrane docking (containing syntaxin 1B/SNAP 23/VAMP 3 <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032603#pone.0032603-Tsai1\" target=\"_blank\">[8]</a>) was not observed in the same fractionated DRM sample at the high M.W. position (at 75 kDa) indicating the dissociation of this SNARE complex upon AE. Arrowheads indicate monomeric form of SNARE proteins present in these fractions. The arrows indicate the expected molecular weight of protein on the Western blot.</p>", "links"=>[], "tags"=>["complexin-snarepin", "non-raft"], "article_id"=>342846, "categories"=>["Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Pei-Shiue J. Tsai", "Ian A. Brewis", "Jillis van Maaren", "Bart M. Gadella"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0032603.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Identification_of_the_complexin_SNAREpin_complex_in_the_non_raft_fractions_/342846", "title"=>"Identification of the complexin-SNAREpin complex in the non-raft fractions.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 08:08:50"}
  • {"files"=>["https://ndownloader.figshare.com/files/672483"], "description"=>"<p>(<b>A</b>) No complexin was recovered on Western blots from membrane cavitates obtained from the non-activated sperm. Complexin emerged at 95 kDa (double arrow heads) in the capacitated membrane cavitates. Complexin showed a calcium-dependent release into a 40 kDa dimer (marked with asterisk) and 20 kDa monomer (marked with single arrow head) as Ca<sup>2+</sup> levels increased. (<b>B</b>) The unexpected stability of the 40 kDa complexin dimer was common to both membrane cavitates and MV samples and showed further dissociation into the expected monomer (arrow head) in the presence of >2 mM Ca<sup>2+</sup> and 1% (v/v) Triton X-100.</p>", "links"=>[], "tags"=>["bicarbonate", "interactions", "complexin"], "article_id"=>342973, "categories"=>["Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Pei-Shiue J. Tsai", "Ian A. Brewis", "Jillis van Maaren", "Bart M. Gadella"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0032603.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Calcium_and_bicarbonate_dependent_interactions_of_complexin_to_protein_complexes_/342973", "title"=>"Calcium and bicarbonate dependent interactions of complexin to protein (complexes).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 08:09:31"}
  • {"files"=>["https://ndownloader.figshare.com/files/672591"], "description"=>"<p>Presented is a subset of the proteins identified for capacitated and acrosome reacted cells, that corresponds to sperm proteins previously reported in the literature to be relevant for sperm-oocyte interactions. Asterisks indicate identified acrosomal membrane proteins, others are plasma membrane specific.</p>*<p>indicates acrosomal specific membrane proteins.</p>", "links"=>[], "tags"=>["ms-based", "proteins", "mvs", "derived", "acrosome", "reacted", "porcine"], "article_id"=>343070, "categories"=>["Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Pei-Shiue J. Tsai", "Ian A. Brewis", "Jillis van Maaren", "Bart M. Gadella"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0032603.t001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_LC_MALDI_MS_based_identification_of_proteins_from_MVs_derived_from_acrosome_reacted_porcine_sperm_/343070", "title"=>"LC-MALDI MS-based identification of proteins from MVs derived from acrosome reacted porcine sperm.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 08:10:03"}
  • {"files"=>["https://ndownloader.figshare.com/files/672630"], "description"=>"<p>Other interesting proteins identified in the MVs derived from bicarbonate capacitated and acrosome reacted cells based on similar criteria to <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032603#pone-0032603-t001\" target=\"_blank\">Table 1</a>. These are proteins that have been previously reported to have roles in secondary sperm-zona binding functioning. Proteins that were not detected in the capacitated membrane cavitates (<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032603#pone-0032603-t003\" target=\"_blank\">Table 3</a>) and are exclusively present in the MVs based on these analyses are highlighted in grey. Most of these proteins have been previously reported to be acrosomal membrane specific <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032603#pone.0032603-VanGestel2\" target=\"_blank\">[29]</a>. Protein ID was considered to be conclusive when two or more peptides were identified (e<0.05) (all proteins except the last protein in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032603#pone-0032603-t002\" target=\"_blank\">Table 2</a>).</p><p>Protein ID was considered conclusive when two or more peptides were identified (e<0.05, all proteins above the line).</p>*<p>: indicate acrosomal specific membrane proteins.</p><p>Highlighted in grey: Proteins that are not detected in the capacitated membrane cavitates and are exclusively present in the mixed vesicles.</p><p>Highlighted in bold: SNARE interacting protein.</p>", "links"=>[], "tags"=>["ms-based", "proteins", "mvs", "derived", "acrosome", "reacted", "porcine"], "article_id"=>343114, "categories"=>["Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Pei-Shiue J. Tsai", "Ian A. Brewis", "Jillis van Maaren", "Bart M. Gadella"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0032603.t002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_LC_MALDI_MS_based_identification_of_other_interesting_proteins_proteins_from_MVs_derived_from_acrosome_reacted_porcine_sperm_/343114", "title"=>"LC-MALDI MS-based identification of other interesting proteins proteins from MVs derived from acrosome reacted porcine sperm.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 08:10:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/672669"], "description"=>"<p>Presented are subsets of those proteins identified that correspond to previously identified sperm proteins relevant to sperm-oocyte interactions <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032603#pone.0032603-VanGestel2\" target=\"_blank\">[29]</a>. SNARE-related protein identifications are highlighted in bold. In addition one proteins with a putative function in the recruitment of SNARE proteins into membrane rafts <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032603#pone.0032603-Ackermann2\" target=\"_blank\">[26]</a> is shown in red. Number signs indicate proteins that have been previously found in membrane cavitates of porcine sperm <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032603#pone.0032603-VanGestel2\" target=\"_blank\">[29]</a>. Asterisks indicate proteins that were identified in the MV preparations (<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032603#pone-0032603-t001\" target=\"_blank\">Tables 1</a> and <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032603#pone-0032603-t002\" target=\"_blank\">2</a>). Protein ID was considered to be conclusive when two or more peptides were identified (e<0.05) (all proteins above the line).</p><p>Protein ID was considered conclusive when two or more peptides were identified (e<0.05, all proteins above the line).</p><p>Highlighted in bold: SNARE proteins or SNARE interacting proteins.</p><p>Highlighted in red: Protein with putative function in the recruitment of SNARE proteins into the membrane raft area.</p>#<p>: indicate proteins which are routinely found in the apical plasma membrane cavitates of porcine sperm.</p>*<p>: indicate proteins that are recovered as plasma membrane proteins in the mixed vesicles.</p>", "links"=>[], "tags"=>["ms-based", "proteins", "membrane", "cavitates", "capacitated", "porcine"], "article_id"=>343152, "categories"=>["Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Pei-Shiue J. Tsai", "Ian A. Brewis", "Jillis van Maaren", "Bart M. Gadella"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0032603.t003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_LC_MALDI_MS_based_identification_of_proteins_from_membrane_cavitates_from_capacitated_porcine_sperm_/343152", "title"=>"LC-MALDI MS-based identification of proteins from membrane cavitates from capacitated porcine sperm.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 08:10:28"}

PMC Usage Stats | Further Information

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Relative Metric

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