Major sperm protein

Last updated
MSP (Major sperm protein) domain
MSPdimer.png
Structure of MSP dimer from A. suum. The β-sheets are shown in orange
Identifiers
SymbolMSP_dom
Pfam PF00635
InterPro IPR000535
PROSITE PS50202
CATH 1MSP
Available protein structures:
Pfam   structures / ECOD  
PDB RCSB PDB; PDBe; PDBj
PDBsum structure summary
Major sperm protein
Identifiers
Organism C. elegans
Symbolmsp-19
Entrez 173981
PDB 1MSP
UniProt P53017
Search for
Structures Swiss-model
Domains InterPro

Major sperm protein (MSP) is a nematode specific small protein of 126 amino acids with a molecular weight of 14 kDa. It is the key player in the motility machinery of nematodes that propels the crawling movement/motility of nematode sperm. It is the most abundant protein present in nematode sperm, comprising 15% of the total protein and more than 40% of the soluble protein. MSP is exclusively synthesized in spermatocytes of the nematodes. [1] The MSP has two main functions in the reproduction of the helminthes: i) as cytosolic component it is responsible for the crawling movement of the mature sperm (without flagellum), and ii) once released, it acts as hormone on the female germ cells, where it triggers oocyte maturation and stimulates the oviduct wall to contract to bring the oocytes into position for fertilization. [2] MSP has first been identified in Caenorhabditis elegans .

Contents

Structure

Molecular structures of MSP from Ascaris suum and Caenorhabditis elegans have been determined by X-ray crystallography [3] and NMR spectroscopy. [4] [5] MSP molecules from these species share 83% sequence identity and their structures are highly similar.

MSP does not harbor any known conserved domain. It is made of a seven-stranded β sandwich, having opposing three-stranded and four-stranded β sheets. Hydrophobic side-chains from adjacent faces in the sandwich form the interior of the protein. The overall structure of MSP resembles an immunoglobulin fold (Ig fold). MSP can be classified as an s-type of this fold, because two of its strands are switching between separate β sheets, unlike in the conserved c-type of the Ig folds. The unique strand switches between the sheets result from two distinct kinks at cis-proline residues 13 and 57 in A. suum protein. [3]

MSP monomers form symmetric dimers. The interaction between MSP monomers in a dimer is very stable, with putative hydrophobic, hydrogen bond and salt bridge interactions. The residues involved in interface formation are between residue 13 and 29 in both A. suum MSP chains of the dimer.

MSP spontaneously polymerises both in vivo and in vitro from dimers into subfilaments, filaments, larger bundles and filament networks. [6]

MSP dimers are the smallest building blocks for these assemblies, none of which have overall polarity:

  1. subfilaments, formed from dimers, connected to a long helix. The dimer-dimer interface within the single subfilament is formed by residues 112-119 of two A. suum MSP chains, which produce an anti-parallel β-strand-β-strand pairing. The interaction is less hydrophobic and results mostly from formation of hydrogen bonds, typically for interfaces between reversibly interacting molecules.
  2. filaments, formed by two subfilaments coiling round one another. The MSP dimer-dimer interactions between two adjacent subfilaments in the filament are characterized by five interfaces, mostly between the residues 78-85 and 98-103. Amino acids 78-85 are part of a highly exposed surface loop connecting different β sheets and are divergent between C. elegans and A. suum. However, the loop consisting of 98-103 residues is highly conserved between all isoforms in both species of the nematode.
  3. fibers, macrofibers or bundles, produced by supercoiling of the filaments. A. suum MSP filaments frequently form rope-like structures called macrofibers. C. elegans MSP mostly form rafts in which a number of filaments are arranged parallel to one another. [4]

In contrast to actin, MSP lacks an ATP-binding site. However, it was noticed that ATP is required for MSP filament assembly at the surface of the plasma membrane. It was suggested that ATP activates either membrane-bound MSP filament end-tracking proteins or their soluble cofactors. [7]

Functions

Sperm motility

Nematode sperm move in an amoeboidal manner by extending a pseudopod. [8] Unlike the motility of actin-based cells, which is based on polar cytoskeletal elements such as actin monomers or tubulin dimers, nematode sperm locomotion is based on a pseudopod and a cytoskeleton built out of a meshwork of non-polar MSP filaments. [1] [9] [10] The two main differences between actin and MSP is that MSP does not bind ATP and the lack of polarity in MSP, thus disabling motility through motor proteins, such as myosin.

Locomotion in nematodes occurs by localized extension of the leading edge of the pseudopod, attachment of the cytoskeleton to the substrate, and retraction of the cell. Assembly of MSP filaments at the leading edge together with disassembly at the base of the pseudopod results in a treadmilling motion, which corresponds to the crawling locomotion of nematode sperm. [11]

Nematode sperm motility is based on a push-pull mechanism which requires two forces triggered by a pH gradient along the pseudopod: one protrusive force and another traction force. The protrusive force is located at the leading edge and pushes against the cell membrane. This force is generated by polymerization of the MSP filaments. MSP filaments are assembled in the cytoplasm near the leading edge of the pseudopod out of MSP dimers resulting in extensions. These extensions allow the interaction of the filament complexes with surrounding complexes, thus resulting in an interconnected uniform cytoskeleton and leading to the crawling movement of the cytoskeleton. MSP filament assembly is triggered by external factors, such as changes in pH, [12] the integral membrane phosphoprotein (MPOP), and MSP domain proteins (MDPs). [11] [13] [14]

A 48 kDa integral membrane phosphoprotein, the major sperm protein polymerization organization protein (MPOP), is the starting point of the pseudopod and is required for the localized membrane-associated polymerization of MSP. [15] This protein is distributed in vesicles throughout the pseudopodal membrane. Tyrosine kinases, which are pH sensitive, phosphorylate the tyrosine residues of MPOP localized at the tip of the pseudopod, thus resulting in the polymerization of the MSP filaments. [16] In Ascaris suum, two MSP-fiber proteins (MFP), MFP1 and 2, with opposite effect on polymerization have been identified. MFP1 inhibits and MFP2 stimulates the MSP assembly. [17] Changes in pH both controls and activates the MSP polymerization throughout spermatogenesis by a pH gradient within the pseudopod of the spermatozoon: assembly occurs at the leading edge where the pH is high, and disassembly of the filaments occurs at the base where the pH is lowered. [11] Degradation of the MSP filaments results in a traction force at the base of the pseudopod, which in turn pulls the cytoskeleton forward. The combination of these two forces is the motive force that allows sperm motility. Attachment of the cytoskeleton to the substratum is required to generate a directional movement. [18] [19]

Affecting female germ cells

MSP affects oocytes on two levels:

  1. MSP regulates oocyte maturation. In C. elegans , oocytes arrest their meiotic cycle at metaphase of meiosis I where it is only resumed in presence of sperms. MSP was identified as the molecular factor triggering oocyte meiotic maturation. It is secreted by the sperms through a vesicular budding mechanism and forms an extracellular concentration gradient. MSP binds to VAB-1 which is an Eph receptor protein-tyrosine kinase on oocytes. In absence of MSP, the VAB-1 Eph receptor inhibits meiotic maturation of oocytes through interaction with inhibitors DAB-1/Disabled and RAN-1. MSP binding prevents this inhibition and results in activation of MAPK pathway. [2] [20] [21] [22]
  2. MSP also stimulates gonadal sheath cell contraction which is a myoepithelial sheath surrounding proximal oocytes. It increases the contraction rate from 10-13 to around 19 contractions per minute. The importance of these contractions is promoting ovulation by inducing the enveloping of the oocyte by the spermatheca. [23]

Homologues

MSP genes have been identified across widely diverged nematode species. They all have more than 60% sequence identity. [1]

Proteins with limited sequence similarity were identified in species from plants to mammals. One of the homologues is VAP33 from Aplysia californica . [24] VAP33 is a protein required for neurotransmitter release, which binds to the v-SNARE synaptobrevin/VAMP, associated with vesicle fusion.

Despite only 11% of sequence similarity, MSP and the N-terminus of the bacterial P-pilus associated chaperonin PapD share a high structural and topological homology in their β sheet regions. Both MSP and PapD can be classified to the s-type immunoglobulin fold proteins, characterized by the above-mentioned unique strand switching. [3]

Related Research Articles

<span class="mw-page-title-main">Spermatozoon</span> Motile sperm cell

A spermatozoon is a motile sperm cell, or moving form of the haploid cell that is the male gamete. A spermatozoon joins an ovum to form a zygote.

<span class="mw-page-title-main">Pseudopodia</span> False leg found on slime molds, archaea, protozoans, leukocytes and certain bacteria

A pseudopod or pseudopodium is a temporary arm-like projection of a eukaryotic cell membrane that is emerged in the direction of movement. Filled with cytoplasm, pseudopodia primarily consist of actin filaments and may also contain microtubules and intermediate filaments. Pseudopods are used for motility and ingestion. They are often found in amoebas.

<span class="mw-page-title-main">Microfilament</span> Filament in the cytoplasm of eukaryotic cells

Microfilaments, also called actin filaments, are protein filaments in the cytoplasm of eukaryotic cells that form part of the cytoskeleton. They are primarily composed of polymers of actin, but are modified by and interact with numerous other proteins in the cell. Microfilaments are usually about 7 nm in diameter and made up of two strands of actin. Microfilament functions include cytokinesis, amoeboid movement, cell motility, changes in cell shape, endocytosis and exocytosis, cell contractility, and mechanical stability. Microfilaments are flexible and relatively strong, resisting buckling by multi-piconewton compressive forces and filament fracture by nanonewton tensile forces. In inducing cell motility, one end of the actin filament elongates while the other end contracts, presumably by myosin II molecular motors. Additionally, they function as part of actomyosin-driven contractile molecular motors, wherein the thin filaments serve as tensile platforms for myosin's ATP-dependent pulling action in muscle contraction and pseudopod advancement. Microfilaments have a tough, flexible framework which helps the cell in movement.

<span class="mw-page-title-main">Epididymis</span> Tube that connects a testicle to a vas deferens

The epididymis is an elongated tubular structure attached to the posterior side of each one of the two male reproductive glands, the testes. It is a single, narrow, tightly coiled tube in adult humans, 6 to 7 meters in length. It connects the testicle to the vas deferens in the male reproductive system. The epididymis serves as an interconnection between the multiple efferent ducts at the rear of a testicle (proximally), and the vas deferens (distally). Its primary function is the storage, maturation and transport of sperm cells.

<span class="mw-page-title-main">Acrosome reaction</span> Sperm-meets-egg process

During fertilization, a sperm must first fuse with the plasma membrane and then penetrate the female egg cell to fertilize it. Fusing to the egg cell usually causes little problem, whereas penetrating through the egg's hard shell or extracellular matrix can be more difficult. Therefore, sperm cells go through a process known as the acrosome reaction, which is the reaction that occurs in the acrosome of the sperm as it approaches the egg.

<span class="mw-page-title-main">Oogenesis</span> Egg cell production process

Oogenesis, ovogenesis, or oögenesis is the differentiation of the ovum into a cell competent to further develop when fertilized. It is developed from the primary oocyte by maturation. Oogenesis is initiated in the embryonic stage.

<span class="mw-page-title-main">Cytokeratin</span>

Cytokeratins are keratin proteins found in the intracytoplasmic cytoskeleton of epithelial tissue. They are an important component of intermediate filaments, which help cells resist mechanical stress. Expression of these cytokeratins within epithelial cells is largely specific to particular organs or tissues. Thus they are used clinically to identify the cell of origin of various human tumors.

<span class="mw-page-title-main">Tropomyosin</span> Protein

Tropomyosin is a two-stranded alpha-helical, coiled coil protein found in many animal and fungal cells. In animals, it is an important component of the muscular system which works in conjunction with troponin to regulate muscle contraction. It is present in smooth and striated muscle tissues, which can be found in various organs and body systems, including the heart, blood vessels, respiratory system, and digestive system. In fungi, tropomyosin is found in cell walls and helps maintain the structural integrity of cells.

<span class="mw-page-title-main">Spermatheca</span> Insect female reproductive organ

The spermatheca, also called receptaculum seminis, is an organ of the female reproductive tract in insects, e.g. ants, bees, some molluscs, oligochaeta worms and certain other invertebrates and vertebrates. Its purpose is to receive and store sperm from the male or, in the case of hermaphrodites, the male component of the body. Spermathecae can sometimes be the site of fertilization when the oocytes are sufficiently developed.

<span class="mw-page-title-main">Sperm</span> Male reproductive cell in anisogamous forms of sexual reproduction

Sperm is the male reproductive cell, or gamete, in anisogamous forms of sexual reproduction. Animals produce motile sperm with a tail known as a flagellum, which are known as spermatozoa, while some red algae and fungi produce non-motile sperm cells, known as spermatia. Flowering plants contain non-motile sperm inside pollen, while some more basal plants like ferns and some gymnosperms have motile sperm.

Cell migration is a central process in the development and maintenance of multicellular organisms. Tissue formation during embryonic development, wound healing and immune responses all require the orchestrated movement of cells in particular directions to specific locations. Cells often migrate in response to specific external signals, including chemical signals and mechanical signals. Errors during this process have serious consequences, including intellectual disability, vascular disease, tumor formation and metastasis. An understanding of the mechanism by which cells migrate may lead to the development of novel therapeutic strategies for controlling, for example, invasive tumour cells.

<span class="mw-page-title-main">Spectrin</span>

Spectrin is a cytoskeletal protein that lines the intracellular side of the plasma membrane in eukaryotic cells. Spectrin forms pentagonal or hexagonal arrangements, forming a scaffold and playing an important role in maintenance of plasma membrane integrity and cytoskeletal structure. The hexagonal arrangements are formed by tetramers of spectrin subunits associating with short actin filaments at either end of the tetramer. These short actin filaments act as junctional complexes allowing the formation of the hexagonal mesh. The protein is named spectrin since it was first isolated as a major protein component of human red blood cells which had been treated with mild detergents; the detergents lysed the cells and the hemoglobin and other cytoplasmic components were washed out. In the light microscope the basic shape of the red blood cell could still be seen as the spectrin-containing submembranous cytoskeleton preserved the shape of the cell in outline. This became known as a red blood cell "ghost" (spectre), and so the major protein of the ghost was named spectrin.

<span class="mw-page-title-main">Synapsis</span> Biological phenomenon in meiosis

Synapsis is the pairing of two chromosomes that occurs during meiosis. It allows matching-up of homologous pairs prior to their segregation, and possible chromosomal crossover between them. Synapsis takes place during prophase I of meiosis. When homologous chromosomes synapse, their ends are first attached to the nuclear envelope. These end-membrane complexes then migrate, assisted by the extranuclear cytoskeleton, until matching ends have been paired. Then the intervening regions of the chromosome are brought together, and may be connected by a protein-RNA complex called the synaptonemal complex. During synapsis, autosomes are held together by the synaptonemal complex along their whole length, whereas for sex chromosomes, this only takes place at one end of each chromosome.

<span class="mw-page-title-main">Cell cortex</span> Layer on the inner face of a cell membrane

The cell cortex, also known as the actin cortex, cortical cytoskeleton or actomyosin cortex, is a specialized layer of cytoplasmic proteins on the inner face of the cell membrane. It functions as a modulator of membrane behavior and cell surface properties. In most eukaryotic cells lacking a cell wall, the cortex is an actin-rich network consisting of F-actin filaments, myosin motors, and actin-binding proteins. The actomyosin cortex is attached to the cell membrane via membrane-anchoring proteins called ERM proteins that plays a central role in cell shape control. The protein constituents of the cortex undergo rapid turnover, making the cortex both mechanically rigid and highly plastic, two properties essential to its function. In most cases, the cortex is in the range of 100 to 1000 nanometers thick.

<span class="mw-page-title-main">Emodepside</span> Chemical compound

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<span class="mw-page-title-main">Amoeboid movement</span> Mode of locomotion in eukaryotic cells

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Oocyteactivation is a series of processes that occur in the oocyte during fertilization.

<span class="mw-page-title-main">Arp2/3 complex</span> Macromolecular complex

Arp2/3 complex is a seven-subunit protein complex that plays a major role in the regulation of the actin cytoskeleton. It is a major component of the actin cytoskeleton and is found in most actin cytoskeleton-containing eukaryotic cells. Two of its subunits, the Actin-Related Proteins ARP2 and ARP3, closely resemble the structure of monomeric actin and serve as nucleation sites for new actin filaments. The complex binds to the sides of existing ("mother") filaments and initiates growth of a new ("daughter") filament at a distinctive 70 degree angle from the mother. Branched actin networks are created as a result of this nucleation of new filaments. The regulation of rearrangements of the actin cytoskeleton is important for processes like cell locomotion, phagocytosis, and intracellular motility of lipid vesicles.

<span class="mw-page-title-main">FERM domain</span>

In molecular biology, the FERM domain is a widespread protein module involved in localising proteins to the plasma membrane. FERM domains are found in a number of cytoskeletal-associated proteins that associate with various proteins at the interface between the plasma membrane and the cytoskeleton. The FERM domain is located at the N terminus in the majority of proteins in which it is found.

<span class="mw-page-title-main">Cortical granule</span>

Cortical granules are regulatory secretory organelles found within oocytes and are most associated with polyspermy prevention after the event of fertilization. Cortical granules are found among all mammals, many vertebrates, and some invertebrates. Within the oocyte, cortical granules are located along the cortex, the region furthest from the cell's center. Following fertilization, a signaling pathway induces the cortical granules to fuse with the oocyte's cell membrane and release their contents into the oocyte's extracellular matrix. This exocytosis of cortical granules is known as the cortical reaction. In mammals, the oocyte's extracellular matrix includes a surrounding layer of perivitelline space, zona pellucida, and finally cumulus cells. Experimental evidence has demonstrated that the released contents of the cortical granules modify the oocyte's extracellular matrix, particularly the zona pellucida. This alteration of the zona pellucida components is known as the zona reaction. The cortical reaction does not occur in all mammals, suggesting the likelihood of other functional purposes for cortical granules. In addition to modifying the oocyte's extracellular matrix and establishing a block to polyspermy, the exocytosis of cortical granules may also contribute towards protection and support of the developing embryo during preimplantation. Once the cortical granules complete their functions, the oocyte does not replenish them.

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