Characteristic properties of proteins from pre-ecdysial cuticle of larvae and pupae of the mealworm Tenebrio molitor.
(2002)
Journal - Insect biochemistry and molecular biology (England )
Abstract :
Proteins extracted from the cuticle of pharate larvae and pupae of the mealworm Tenebrio molitor are more soluble at low temperatures than at higher temperatures, a behaviour characteristic of hydrophobic proteins. When the temperature of an unfractionated cuticular extract is raised from 4 to 25 degrees C the solution becomes turbid, droplets of a heavy, protein-rich phase are formed, which gradually settles, leaving an upper protein-poor phase, indicating that the aggregation process is a coacervation. The aggregation of the dissolved cuticular proteins is influenced by changes in temperature, pH, and ionic strength. The process has been studied by measuring development of turbidity in unfractionated cuticular extracts and in solutions of three purified proteins from Tenebrio pharate larvae and pupae (TmLPCP-A1a, TmLPCP-E1a, and TmLPCP-G1a), while temperature, pH or ionic strength of the solutions were varied. Protein aggregation was also studied by determination of changes in fluorescence intensity, when the hydrophobicity probe, 8-anilinonaphthalenesulfonic acid (ANS) was added to solutions of the cuticular proteins. Only when the protein solutions had developed a measurable turbidity was an increase in ANS-fluorescence observed, indicating formation of tightly packed clusters of hydrophobic amino acid residues during aggregation. The temperature range for aggregation depends upon protein concentration: the higher the concentration the lower and more narrow is the temperature range within which aggregation occurs. The tendency for the individual cuticular proteins to aggregate is most pronounced near their isoelectric points, and most of the cuticular proteins have alkaline isoelectric points. The influence of salts on the tendency of the proteins to aggregate varies among the proteins and depends upon how close they are to their isoelectric point. A solution containing both protein TmLPCP-A1a and TmLPCP-E1a becomes more turbid and develops a more intense ANS-fluorescence when warmed from 10 to 30 degrees C than corresponding to the sum of measurements performed on separate solutions of the two proteins, indicating that the two proteins interact during aggregation. The Tenebrio larval/pupal cuticular proteins are characterized by an abundance of hydrophobic amino acid residues, and especially their contents of alanine and proline are high. The behaviour of the cuticular proteins in solution resembles that of another hydrophobic protein, tropoelastin, and it seems reasonable to suggest that similar interactions govern the folding and aggregation of the peptide chains in the two types of proteins. The proline and alanine rich chain segments in the pharate cuticular proteins are suggested to form a series of beta-turns and to fold into a relatively open structure at low temperatures, giving water access to the hydrophobic residues and making the proteins water soluble. At increased temperatures the structure of the ordered water layer surrounding the hydrophobic groups breaks down, and the peptide chains tend to collapse into a more closed structure and to interact more tightly with hydrophobic regions in neighbouring molecules. In dilute solutions in the test tube this results in aggregation and precipitation of the proteins; in intact, pharate cuticle at ambient temperatures the proteins will preferably be in an aggregated, easily dissociated state. Accordingly, small changes in intercuticular pH and ionic strength can produce pronounced changes in the mechanical properties of unsclerotized solid cuticle by interference with protein interactions, in agreement with reports that some cuticles undergo plasticization during and/or immediately after ecdysis.
| ISSN : | 0965-1748 |
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| Mesh Heading : | Animals Hydrogen-Ion Concentration Insect Proteins Larva Osmolar Concentration Pupa Tenebrio |
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| Mesh Heading Relevant : | analysis chemistry |
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Sequence studies of proteins from larval and pupal cuticle of the yellow meal worm, Tenebrio molitor.
(1997)
Journal - Insect biochemistry and molecular biology (ENGLAND )
Abstract :
Complete amino acid sequences have been determined for six larval-pupal cuticular proteins from Tenebrio molitor. The sequenced proteins are major components in both larval and pupal cuticle, and both basic and slightly acidic proteins are represented. The proteins show pronounced similarities to some of the proteins sequenced from other insect cuticles. Three slightly acidic larval-pupal Tenebrio cuticular proteins contain a 66-residue central, hydrophilic region, resembling regions in cuticular proteins from insect species of four different orders (Coleoptera, Diptera, Lepidoptera and Orthoptera), and three basic proteins from larval-pupal Tenebrio cuticle have a 51-residue hydrophilic region in common with two proteins from cuticle of pharate adult locusts (Locusta migratoria). The Tenebrio larval-pupal cuticular proteins are also similar to locust adult cuticular proteins, by frequent occurrence of the short sequence motif Ala-Ala-Pro-Ala/Val. The pronounced sequence similarities between cuticular proteins from different insect orders indicate that the conserved regions are functionally important.
| ISSN : | 0965-1748 |
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| Mesh Heading : | Amino Acid Sequence Animals Electrophoresis, Gel, Two-Dimensional Insect Proteins Larva Molecular Sequence Data Pupa Sequence Homology, Amino Acid Tenebrio |
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| Mesh Heading Relevant : | chemistry chemistry |
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Insect cuticular proteins.
(1995)
Journal - Insect biochemistry and molecular biology (ENGLAND )
Abstract :
Insect cuticles are composite structural materials with mechanical properties optimal for their biological functions. The bulk properties of cuticles are to a large extent determined by the interactions between the various components, mainly the chitin filament system and the proteins. The various cuticular types show pronounced differences in mechanical properties, and it is suggested that these differences can be related to the properties of the individual proteins and to the degree of secondary stabilization (sclerotization). The amino acid sequences, which have been obtained for insect cuticular proteins either by direct sequencing of purified proteins or by deduction from corresponding DNA-sequences, are listed according to insect order and species. Extensive sequence similarity is observed among several cuticular proteins obtained from different insect orders. Other cuticular proteins are characterized by repeated occurrence of a few small motifs consisting mainly of hydrophobic residues. The latter group of proteins has so far only been reported from stiff cuticles. The possible relevance of the various motifs and repeats for protein interaction and the mechanical properties of cuticles is discussed.
| ISSN : | 0965-1748 |
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| Mesh Heading : | Amino Acid Sequence Animals Molecular Sequence Data Protein Conformation Sequence Homology, Amino Acid analysis classification |
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| Mesh Heading Relevant : | Insect Proteins Insects Proteins |
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Comparison of larval and pupal cuticular proteins in Tenebrio molitor.
(1995)
Journal - Insect biochemistry and molecular biology (ENGLAND )
Abstract :
Protein extracts from pupal and larval pharate cuticle from the meal worm, Tenebrio molitor, gave nearly identical patterns by two-dimensional electrophoresis and by ion-exchange chromatography. The main components in the cuticular extracts from the two metamorphic stages were also identical with respect to molecular mass according to electrospray ionization mass spectrometry. The complete amino acid sequence for one of the pupal cuticular proteins was determined; according to partial amino acid sequences and the mass spectrometric peptide map for the corresponding larval cuticular protein, it was concluded that the larval protein has the same amino acid sequence as the pupal protein. The sequence is characterized by a high content of alanine, proline, valine, and tyrosine and the complete absence of acidic amino acid residues, the sulphur containing amino acids and tryptophan. The sequence is further characterized by a high frequency of repeated sequence motifs, among which the Ala-Ala-Pro-Ala motif is the most abundant, but also longer sequence motifs are repeated. The sequence shows striking resemblance to sequences of proteins isolated from pharate locust cuticle.
| ISSN : | 0965-1748 |
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| Mesh Heading : | Amino Acid Sequence Animals Larva Molecular Sequence Data Peptide Fragments Proteins Pupa Sequence Analysis Tenebrio chemistry analysis isolation & purification chemistry |
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| Mesh Heading Relevant : | Insect Proteins chemistry chemistry |
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