SpermatogenesisOnline 1.0

Tag Content
SG ID
SG00019123 
UniProt Accession
Theoretical PI
6.36  
Molecular Weight
47141 Da  
Genbank Nucleotide ID
Genbank Protein ID
Gene Name
Eno1 
Gene Synonyms/Alias
Eno-1 
Protein Name
Alpha-enolase 
Protein Synonyms/Alias
EC=4.2.1.11 2-phospho-D-glycerate hydro-lyase; Enolase 1; Non-neural enolase;NNE 
Organism
Mus musculus (Mouse) 
NCBI Taxonomy ID
10090 
Chromosome Location
chr:18;48204989-48208032;1
View in Ensembl genome browser  
Function in Stage
Uncertain 
Function in Cell Type
Uncertain 
Probability (GAS) of Function in Spermatogenesis
0.01313815 
The probability was calculated by GAS algorithm, ranging from 0 to 1. The closer it is to 1, the more possibly it functions in spermatogenesis.
Description
Temporarily unavailable 
Abstract of related literatures
2. This study describes comprehensive polling of transcription start and termination sites and analysis of previously unidentified full-length complementary DNAs derived from the mouse genome. We identify the 5' and 3' boundaries of 181,047 transcripts with extensive variation in transcripts arising from alternative promoter usage, splicing, and polyadenylation. There are 16,247 new mouse protein-coding transcripts, including 5154 encoding previously unidentified proteins. Genomic mapping of the transcriptome reveals transcriptional forests, with overlapping transcription on both strands, separated by deserts in which few transcripts are observed. The data provide a comprehensive platform for the comparative analysis of mammalian transcriptional regulation in differentiation and development. PMID: [16141072] 

3. The National Institutes of Health's Mammalian Gene Collection (MGC) project was designed to generate and sequence a publicly accessible cDNA resource containing a complete open reading frame (ORF) for every human and mouse gene. The project initially used a random strategy to select clones from a large number of cDNA libraries from diverse tissues. Candidate clones were chosen based on 5'-EST sequences, and then fully sequenced to high accuracy and analyzed by algorithms developed for this project. Currently, more than 11,000 human and 10,000 mouse genes are represented in MGC by at least one clone with a full ORF. The random selection approach is now reaching a saturation point, and a transition to protocols targeted at the missing transcripts is now required to complete the mouse and human collections. Comparison of the sequence of the MGC clones to reference genome sequences reveals that most cDNA clones are of very high sequence quality, although it is likely that some cDNAs may carry missense variants as a consequence of experimental artifact, such as PCR, cloning, or reverse transcriptase errors. Recently, a rat cDNA component was added to the project, and ongoing frog (Xenopus) and zebrafish (Danio) cDNA projects were expanded to take advantage of the high-throughput MGC pipeline. PMID: [15489334] 

4. This report explores the hypothesis that massive cholesteryl ester (CE) accumulation in macrophages, such as that occurring in atheroma foam cells, results in changes in the expression or modification of specific cellular proteins. Two-dimensional (2-D) gel electrophoretic patterns of metabolically labeled cellular proteins from mouse peritoneal macrophages that were loaded with CE (through incubation with acetylated low density lipoprotein [acetyl-LDL] for 4 days) were compared with those of control macrophages. Densitometric analysis of 2-D gel autoradiograms from the cell lysates revealed statistically significant changes in seven cellular proteins (five decreases and two increases). The changes in protein expression (foam cell versus control) ranged from a 458 +/- 164% (p < 0.001) increase to a 35 +/- 34% (p < 0.001) decrease (n = 11). Incubation of macrophages with beta-very low density lipoprotein, which also increased the CE content of macrophages (albeit to a lesser extent than acetyl-LDL), resulted in changes in five of the seven proteins. In contrast, incubation of cells with LDL, fucoidan, or latex beads, none of which caused CE accumulation, did not lead to significant changes in four of these five proteins. One of these four proteins, which increased fourfold to fivefold in foam cells (M(r) = 49,000; isoelectric point of 6.8), was purified by preparative 2-D gel electrophoresis. Internal amino acid sequence of cyanogen bromide fragments of this protein as well as Western blot analysis identified this protein as an isoform of alpha-enolase. The increased expression of this alpha-enolase isoform, which was seen as early as day 2 of acetyl-LDL incubation of the macrophages, was diminished by including an inhibitor of cholesterol esterification during the acetyl-LDL incubation period. In conclusion, macrophage foam cell formation is associated with distinct changes in protein expression, including a marked increase in an isoform of alpha-enolase, suggesting a specific biological adaptation to CE loading. PMID: [8427861] 

5. The glycolytic enzyme enolase (EC 4.2.1.11) is active as dimers formed from three subunits encoded by different genes. The embryonic alphaalpha isoform remains distributed in many adult cell types, whereas a transition towards betabeta and gammagamma isoforms occurs in striated muscle cells and neurons respectively. It is not understood why enolase exhibits tissue-specific isoforms with very close functional properties. We approached this problem by the purification of native betabeta-enolase from mouse hindlimb muscles and by raising specific antibodies of high titre against this protein. These reagents have been useful in revealing a heterogeneity of the beta-enolase subunit that changes with in vivo and in vitro maturation. A basic carboxypeptidase appears to be involved in generating an acidic beta-enolase variant, and may regulate plasminogen binding by this subunit. We show for the first time that pure betabeta-enolase binds with high affinity the adjacent enzymes in the glycolytic pathway (pyruvate kinase and phosphoglycerate mutase), favouring the hypothesis that these three enzymes form a functional glycolytic segment. betabeta-Enolase binds with high affinity sarcomeric troponin but not actin and tropomyosin. Some of these binding properties are shared by the alphaalpha-isoenolase, which is also expressed in striated muscle, but not by the neuron-specific gammagamma-enolase. These results support the idea that specific interactions with macromolecules will address muscle enolase isoforms at the subcellular site where ATP, produced through glycolysis, is most needed for contraction. Such a specific targeting could be modulated by post-translational modifications. PMID: [9169614] 

6. Enolase is a dimeric glycolytic enzyme exhibiting tissue specific isoforms. During ontogenesis, a transition occurs from the embryonic alphaalpha towards the specific alphabeta, and betabeta isoforms in striated muscle. Immunocytochemical analyses on transverse sections of adult mouse gastrocnemius muscle, allowed us to compare the expression of alpha and beta subunits to that of myosin heavy chain (MHC) isoforms. Levels of beta immunoreactivity followed the order IIB > IIX > IIA > I. This gradient parallels the ATPase activity associated to MHC isoforms, indicating that the expression of beta enolase in myofibres is finely regulated as a function of energetic requirements. By contrast, variations in alpha immunolabelling intensity appeared independent of fibre types. Longitudinal muscle sections exhibited a striated pattern of alpha immunoreactivity. Confocal microscopy analyses demonstrated that alpha was localised at the M band. Most beta immunoreactivity was diffuse all over the sarcoplasm. However, some beta immunoreactivity was striated and localized at both Z and M bands. Thus, betabeta enolase could participate to multi-enzyme complexes present at the I band, and involved with local ATP production. Our results support the notion that isozymes differ in their ability to interact with other macromolecules, thus segregating to different subcellular sites where they would respond to specific functional demands. PMID: [11229603] 

7. Though the interferon-inducible protein ISG15 was one of the first ubiquitin-like modifiers to be discovered, much remains unknown about the identity of proteins conjugated to ISG15 or the biologic consequences of modification. To gain a better understanding of the cellular pathways affected by ISG15, we identified proteins targeted for ISGylation using a proteomic approach. Mass spectrometric analysis identified 76 candidate ISGylation targets in anti-ISG15 immunoprecipitates from interferon-treated mouse or human cells. Twenty-one proteins were found in both mouse and human samples, including STAT1, a known target of ISGylation. Candidates identified in both species were tested for ISGylation in a transfection system: 18 of 19 proteins tested were ISGylated in this system. Two candidates, EF-2 and VCP, were also shown to be ISGylated in an interferon-dependent manner in the absence of exogenous over-expression. Seven proteins identified from a single species, but functionally related to candidates found in both species, were also ISGylated in the over-expression system. Proteins that can be ISGylated play important roles in translation, glycolysis, stress responses, and cell motility. These data indicate that ISGylation targets proteins found in several fundamentally important cellular pathways and will contribute to understanding the physiologic role of interferon-induced ISG15 and ISG15 conjugation. PMID: [16139798] 

8. Metazoans employ reversible tyrosine phosphorylation to regulate innumerable biological processes. Thus, the large-scale identification of tyrosine phosphorylation sites from primary tissues is an essential step toward a molecular systems understanding of dynamic regulation in vivo. The relative paucity of phosphotyrosine has greatly limited its identification in large-scale phosphoproteomic experiments. However, using antiphosphotyrosine peptide immunoprecipitations, we report the largest study to date of tyrosine phosphorylation sites from primary tissue, identifying 414 unique tyrosine phosphorylation sites from murine brain. To measure the conservation of phosphorylated tyrosines and their surrounding residues, we constructed a computational pipeline and identified patterns of conservation within the signature of phosphotyrosine. PMID: [18034455] 

9. Activity-dependent protein phosphorylation is a highly dynamic yet tightly regulated process essential for cellular signaling. Although recognized as critical for neuronal functions, the extent and stoichiometry of phosphorylation in brain cells remain undetermined. In this study, we resolved activity-dependent changes in phosphorylation stoichiometry at specific sites in distinct subcellular compartments of brain cells. Following highly sensitive phosphopeptide enrichment using immobilized metal affinity chromatography and mass spectrometry, we isolated and identified 974 unique phosphorylation sites on 499 proteins, many of which are novel. To further explore the significance of specific phosphorylation sites, we used isobaric peptide labels and determined the absolute quantity of both phosphorylated and non-phosphorylated peptides of candidate phosphoproteins and estimated phosphorylation stoichiometry. The analyses of phosphorylation dynamics using differentially stimulated synaptic terminal preparations revealed activity-dependent changes in phosphorylation stoichiometry of target proteins. Using this method, we were able to differentiate between distinct isoforms of Ca2+/calmodulin-dependent protein kinase (CaMKII) and identify a novel activity-regulated phosphorylation site on the glutamate receptor subunit GluR1. Together these data illustrate that mass spectrometry-based methods can be used to determine activity-dependent changes in phosphorylation stoichiometry on candidate phosphopeptides following large scale phosphoproteome analysis of brain tissue. PMID: [17114649] 

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Function
Multifunctional enzyme that, as well as its role inglycolysis, plays a part in various processes such as growthcontrol, hypoxia tolerance and allergic responses (By similarity).May also function in the intravascular and pericellularfibrinolytic system due to its ability to serve as a receptor andactivator of plasminogen on the cell surface of several cell-typessuch as leukocytes and neurons. Stimulates immunoglobulinproduction (By similarity). 
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Subcellular Location
Cytoplasm (By similarity). Cell membrane (Bysimilarity). Note=Can translocate to the plasma membrane in eitherthe homodimeric (alpha/alpha) or heterodimeric (alpha/gamma) form(By similarity). ENO1 is localized to the M-band. 
Tissue Specificity
The alpha/alpha homodimer is expressed inembryo and in most adult tissues. The alpha/beta heterodimer andthe beta/beta homodimer are found in striated muscle, and thealpha/gamma heterodimer and the gamma/gamma homodimer in neurons.In striated muscle, expression of ENO1 appears to be independentof fiber type. 
Gene Ontology
GO IDGO termEvidence
GO:0000015 C:phosphopyruvate hydratase complex IEA:InterPro.
GO:0005886 C:plasma membrane IEA:UniProtKB-SubCell.
GO:0000287 F:magnesium ion binding IEA:InterPro.
GO:0004634 F:phosphopyruvate hydratase activity IEA:EC.
GO:0006096 P:glycolysis IEA:UniProtKB-KW.
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Interpro
IPR000941;    Enolase.
IPR020810;    Enolase_C.
IPR020809;    Enolase_CS.
IPR020811;    Enolase_N.
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Pfam
PF00113;    Enolase_C;    1.
PF03952;    Enolase_N;    1.
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SMART
PROSITE
PS00164;    ENOLASE;    1.
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PRINTS
PR00148;    ENOLASE.;   
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Created Date
18-Oct-2012 
Record Type
GAS predicted 
Sequence Annotation
INIT_MET      1      1       Removed.
CHAIN         2    434       Alpha-enolase.
                             /FTId=PRO_0000134098.
REGION      370    373       Substrate binding (By similarity).
REGION      405    434       Required for interaction with PLG (By
                             similarity).
ACT_SITE    210    210       Proton donor (By similarity).
ACT_SITE    343    343       Proton acceptor (By similarity).
METAL       245    245       Magnesium (By similarity).
METAL       293    293       Magnesium (By similarity).
METAL       318    318       Magnesium (By similarity).
BINDING     158    158       Substrate (By similarity).
BINDING     167    167       Substrate (By similarity).
BINDING     293    293       Substrate (By similarity).
BINDING     318    318       Substrate (By similarity).
BINDING     394    394       Substrate (By similarity).
MOD_RES       2      2       N-acetylserine (By similarity).
MOD_RES      44     44       Phosphotyrosine.
MOD_RES      60     60       N6-acetyllysine (By similarity).
MOD_RES      63     63       Phosphoserine (By similarity).
MOD_RES      71     71       N6-acetyllysine (By similarity).
MOD_RES      72     72       Phosphothreonine (By similarity).
MOD_RES      89     89       N6-acetyllysine (By similarity).
MOD_RES     126    126       N6-acetyllysine (By similarity).
MOD_RES     193    193       N6-acetyllysine (By similarity).
MOD_RES     199    199       N6-acetyllysine (By similarity).
MOD_RES     228    228       N6-acetyllysine (By similarity).
MOD_RES     233    233       N6-acetyllysine; alternate (By
                             similarity).
MOD_RES     233    233       N6-malonyllysine; alternate (By
                             similarity).
MOD_RES     254    254       Phosphoserine (By similarity).
MOD_RES     256    256       N6-acetyllysine (By similarity).
MOD_RES     263    263       Phosphoserine.
MOD_RES     281    281       N6-acetyllysine (By similarity).
MOD_RES     287    287       Phosphotyrosine (By similarity).
MOD_RES     406    406       N6-acetyllysine (By similarity).
MOD_RES     420    420       N6-acetyllysine; alternate (By
                             similarity).
MOD_RES     420    420       N6-malonyllysine; alternate (By
                             similarity).
CONFLICT    359    359       L -> P (in Ref. 1; CAA36605).
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Nucleotide Sequence
Length: 1720 bp   Go to nucleotide: FASTA
Protein Sequence
Length: 434 bp   Go to amino acid: FASTA
The verified Protein-Protein interaction information
UniProt
 Gene Symbol Ref Databases
YwhabIntAct 
ACHEIntAct 
Kcnma1IntAct 
PrkceIntAct 
SETIntAct 
Traf6BioGRID 
Other Protein-Protein interaction resources
String database  
View Microarray data
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