共查询到20条相似文献,搜索用时 46 毫秒
1.
2.
3.
A group of related cDNAs encoding secreted proteins from Hessian fly [Mayetiola destructor (Say)] salivary glands 总被引:2,自引:0,他引:2
A group of cDNAs has been isolated and characterized from Hessian fly [Mayetiola destructor (Say)] salivary glands. Members in this group appear to encode proteins with secretion signal peptides at the N-terminals. The mature putative proteins are small, basic proteins with calculated molecular weights that ranged from 8.5 to 10 kDa, and isoelectric points from 9.92 to 10.90. Sequence analysis indicated a strong selection for mutations that generate amino acid changes within the coding region. Northern blot analysis revealed that these genes are expressed only in the first instar larvae, a critical stage that determines if the interaction between a specific Hessian fly biotype and a specific wheat cultivar is compatible. Genomic analysis demonstrated that multiple copies of similar genes are clustered within a short region on chromosome 2A. This is the same arm in which two avirulence genes have been mapped. 相似文献
4.
Differential expression of candidate salivary effector proteins in field collections of Hessian fly,Mayetiola destructor
下载免费PDF全文
![点击此处可从《Insect molecular biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
A. J. Johnson R. H. Shukle M.‐S. Chen S. Srivastava S. Subramanyam B. J. Schemerhorn P. G. Weintraub H. E. M. Abdel Moniem K. L. Flanders G. D. Buntin C. E. Williams 《Insect molecular biology》2015,24(2):191-202
5.
Isolation and sequence analysis of serine protease cDNAs from mouse cytolytic T lymphocytes 总被引:3,自引:5,他引:3
下载免费PDF全文
![点击此处可从《The Journal of experimental medicine》网站下载免费的PDF全文](/ch/ext_images/free.gif)
《The Journal of experimental medicine》1988,168(5):1839-1854
Three new cDNA clones (designated MCSP-1, MCSP-2, and MCSP-3) encoding mouse serine proteases were isolated from cloned cytolytic T lymphocytes (CTL) by a modified differential screening procedure. The putative mature proteins of MCSP-2 and MCSP-3 are each composed of 228 amino acids with molecular weights of 25,477 and 25,360, respectively. NH2-terminal amino acids of MCSP-2- and MCSP-3-predicted proteins were identical to those reported for granzyme E and F, respectively. The third species, MCSP-1, was closely related to the two other cDNA species but approximately 30 amino acids equivalents of the NH2- terminal portion of the cDNA were not cloned. The amino acids forming the active sites of serine proteases were well conserved among the three predicted proteins. The active site pocket residue positioned six residues before the active-site Ser184 is alanine in MCSP-1, threonine in MCSP-2, and serine in MCSP-3, indicating that both MCSP-2 and MCSP-3 may have chymotrypsin-like specificity. There are three potential asparagine-linked glycosylation sites in MCSP-1 and MCSP-3, and four in MCSP-2-deduced amino acid sequences. Amino acid comparison of MCSP-1 with four other reported serine proteases whose active site pocket residue is alanine revealed that MCSP-1 was substantially different from the other molecules, indicating that MCSP-1 may be a new member of mouse T cell serine protease family. Antibodies made against a MCSP-1 lacZ gene fusion protein stain granules of CTL and react on immunoblots with two distinct granule protein bands of 29 and 35-40 kD. Only the 35- kD species labels with [3H]DFP. Since a protease cascade may play a key role in cytolytic lymphocyte activation, our isolation of cDNAs representative of unique serine esterases should help to investigate such a cascade process. 相似文献
6.
7.
The Hessian fly (Mayetiola destructor) is an agriculturally important pest of wheat. A mariner element (Desmar1) has been previously identified in the Hessian fly genome. Using Desmar1 as a probe, we isolated individual copies of Desmar‐like elements from the Hessian fly genome cloned in bacterial artificial chromosomes (BACs) and studied their structural variability and flanking DNA sequences. The partial Desmar‐like copies are relatively more abundant (~64%) than full length copies (~36%) in the Hessian fly genome. Most of the full length copies are consistently flanked by an EcoRI restriction site that occurs 32 bp from one end and 66 bp from the other end of the mariner. Using an amplified fragment length polymorphism‐PCR (AFLP‐PCR) based method, we identified segregating polymorphisms associated with Desmar elements in a F2 mapping population. We were able to use the segregation data to localize the chromosomal position of three Desmar elements by linkage analysis. As paternal chromosomes are eliminated in the Hessian fly during early embryogenesis, two‐thirds of the AFLPs were expected to be polymorphic in the mapping population and this was observed for AFLPs anchored to full length Desmar copies but not to the partial copies. Thus, our data indicate that dead and partial Desmar‐like copies are probably associated with less polymorphic regions and may represent mariner graveyards in the Hessian fly genome. 相似文献
8.
Cloning of a family of serine protease genes from the cat flea Ctenocephalides felis 总被引:1,自引:0,他引:1
Serine protease gene fragments approximately 480 nucleotides in length were amplified from Ctenocephalides felis larval and adult cDNA libraries using degenerate oligonucleotide PCR primers. Partial clones of thirty-eight distinct serine protease encoding sequences were isolated, and nineteen different full-length cDNAs encoding mature serine proteases were subsequently cloned and sequenced. All of the mature proteases contained the histidine, aspartic acid and serine amino acids of the catalytic triad characteristic of serine proteases. The mature C. felis serine proteases had amino acid sequences that were at most 29–53% identical to those known insect and arachnid serine proteases. Two of the C. felis gene sequences had similarity with the Drosophila melanogaster developmental genes snake and stubble. mRNA expression of selected serine protease genes was examined in different life stages, tissues, genders, and in response to bloodfeeding. 相似文献
9.
Ole E. S?rensen Stine N. Clemmensen Sara L. Dahl Ole ?stergaard Niels H. Heegaard Andreas Glenth?j Finn Cilius Nielsen Niels Borregaard 《The Journal of clinical investigation》2014,124(10):4539-4548
Papillon-Lefèvre syndrome (PLS) results from mutations that inactivate cysteine
protease cathepsin C (CTSC), which processes a variety of serine proteases considered
essential for antimicrobial defense. Despite serine protease–deficient immune cell
populations, PLS patients do not exhibit marked immunodeficiency. Here, we characterized a
24-year-old woman who had suffered from severe juvenile periodontal disease, but was
otherwise healthy, and identified a homozygous missense mutation in CTSC
indicative of PLS. Proteome analysis of patient neutrophil granules revealed that several
proteins that normally localize to azurophil granules, including the major serine
proteases, elastase, cathepsin G, and proteinase 3, were absent. Accordingly, neutrophils
from this patient were incapable of producing neutrophil extracellular traps (NETs) in
response to ROS and were unable to process endogenous cathelicidin hCAP-18 into the
antibacterial peptide LL-37 in response to ionomycin. In immature myeloid cells from
patient bone marrow, biosynthesis of CTSC and neutrophil serine proteases appeared normal
along with initial processing and sorting to cellular storage. In contrast, these proteins
were completely absent in mature neutrophils, indicating that CTSC
mutation promotes protease degradation in more mature hematopoietic subsets, but does not
affect protease production in progenitor cells. Together, these data indicate CTSC
protects serine proteases from degradation in mature immune cells and suggest that
neutrophil serine proteases are dispensable for human immunoprotection. 相似文献
10.
Y. R. Chikate V. A. Tamhane R. S. Joshi V. S. Gupta A. P. Giri 《Insect molecular biology》2013,22(3):258-272
Helicoverpa armigera (Lepidoptera: Noctuidae) and other polyphagous agricultural pests are extending their plant host range and emerging as serious agents in restraining crop productivity. Dynamic regulation, coupled with a diversity of digestive and detoxifying enzymes, play a crucial role in the adaptation of polyphagous insects. To investigate the functional intricacy of serine proteases in the development and polyphagy of H. armigera, we profiled the expression of eight trypsin‐like and four chymotrypsin‐like phylogenetically diverse mRNAs from different life stages of H. armigera reared on nutritionally distinct host plants. These analyses revealed diet‐ and stage‐specific protease expression patterns. The trypsins expressed showed structural variations, which might result in differential substrate specificity and interaction with inhibitors. Protease profiles in the presence of inhibitors and their mass spectrometric analyses revealed insight into their differential activity. These findings emphasize the differential expression of serine proteases and their consequences for digestive physiology in promoting polyphagy in H. armigera. 相似文献
11.
The structure and mechanism of bacterial type I signal peptidases. A novel antibiotic target 总被引:7,自引:0,他引:7
Type I signal peptidases are essential membrane-bound serine proteases that function to cleave the amino-terminal signal peptide extension from proteins that are translocated across biological membranes. The bacterial signal peptidases are unique serine proteases that utilize a Ser/Lys catalytic dyad mechanism in place of the classical Ser/His/Asp catalytic triad mechanism. They represent a potential novel antibiotic target at the bacterial membrane surface. This review will discuss the bacterial signal peptidases that have been characterized to date, as well as putative signal peptidase sequences that have been recognized via bacterial genome sequencing. We review the investigations into the mechanism of Escherichia coli and Bacillus subtilis signal peptidase, and discuss the results in light of the recent crystal structure of the E. coli signal peptidase in complex with a beta-lactam-type inhibitor. The proposed conserved structural features of Type I signal peptidases give additional insight into the mechanism of this unique enzyme. 相似文献
12.
13.
Simpson RM Newcomb RD Gatehouse HS Crowhurst RN Chagné D Gatehouse LN Markwick NP Beuning LL Murray C Marshall SD Yauk YK Nain B Wang YY Gleave AP Christeller JT 《Insect molecular biology》2007,16(6):675-690
The midgut is a key tissue in insect science. Physiological roles include digestion and peritrophic membrane function, as well as being an important target for insecticides. We used an expressed sequence tag (EST) approach to identify candidate genes and gene families involved in these processes in the light brown apple moth, Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae). Two cDNA libraries were constructed from dissected midgut of third to fifth instar larvae. Clustering analysis of 6416 expressed sequence tags produced 1178 tentative unique genes comprising 725 tentative contigs and 453 singletons. The sequences show similar codon usage to sequences from other lepidopterans, a Kozak consensus sequence similar to Drosophila and single nucleotide polymorphisms (SNPs) were detected at a frequency of 1.35/kb. The identity of the most common Interpro families correlates well with major known functions of the midgut. Phylogenetic analysis was conducted on representative sequences from selected multigene families. Gene families include a broad range of digestive proteases, lipases and carbohydrases that appear to have degradative capacity against the major food components found in leaves, the diet of these larvae; and carboxylesterases, glutathione-S-transferases and cytochrome P450 monooxygenases, potentially involved in xenobiotic degradation. Two of the larger multigene families, serine proteases and lipases, expressed a high proportion of genes that are likely to be catalytically inactive. 相似文献
14.
C. L. Russell K. J. Heesom† C. J. Arthur‡ C. R. Helps P. S. Mellor§ M. J. Day S. Torsteinsdottir¶ T. S. Björnsdóttir¶ A. D. Wilson 《Insect molecular biology》2009,18(3):383-393
Culicoides spp. are vectors of several infectious diseases of veterinary importance and a major cause of allergy in horses and other livestock. Their saliva contains a number of proteins which enable blood feeding, enhance disease transmission and act as allergens. We report the construction of a novel cDNA library from Culicoides nubeculosus linked to the analysis of abundant salivary gland proteins by mass spectrometry. Fifty-four novel proteins sequences are described including those of the enzymes maltase, hyaluronidase and two serine proteases demonstrated to be present in Culicoides salivary glands, as well as several members of the D7 family and protease inhibitors with putative anticoagulant activity. In addition, several families of abundant proteins with unknown function were identified including some of the major candidate allergens that cause insect bite hypersensitivity in horses. 相似文献
15.
Marshall SD Gatehouse LN Becher SA Christeller JT Gatehouse HS Hurst MR Boucias DG Jackson TA 《Insect molecular biology》2008,17(3):247-259
Costelytra zealandica larvae are pests of New Zealand pastures causing damage by feeding on the roots of grasses and clovers. The major larval protein digestive enzymes are serine proteases (SPs), which are targets for disruption in pest control. An expressed sequence tag (EST) library from healthy, third instar larval midgut tissue was constructed and analysed to determine the composition and regulation of proteases in the C. zealandica larval midgut. Gene mining identified three trypsin-like and 11 chymotrypsin-like SPs spread among four major subgroups. Representative SPs were examined by quantitative PCR and enzyme activity assayed across developmental stages. The serine protease genes examined were expressed throughout feeding stages and downregulated in nonfeeding stages. The study will improve targeting of protease inhibitors and bacterial disruptors of SP synthesis. 相似文献
16.
17.
C. M. Elvin T. Vuocolo W. J. M. Smith C. H. Eisemann P. W. Riddles 《Insect molecular biology》1994,3(2):105-115
A large and diverse family of serine protease genes was identified in first-instar larval cDNA of the sheep blowfly (Lucilia cuprina). This complex repertoire of genes was identified via a PCR approach using highly degenerate primers based on structurally conserved regions which surround the active site His and Ser residues found in all serine proteases. PCR products from entire first-instar larval cDNA, or from third-instar larval salivary glands or cardia, generated using a microscale RT-PCR method, were cloned into a plas-mid vector. Comparison of the restriction fragment patterns of PCR products generated from the three different sources suggests a highly diverse tissue-specific pattern of serine protease expression in this organism. Detailed analysis of the restriction fragment patterns of sixty-nine randomly selected clones from entire first-instar larvae revealed forty-nine different classes of PCR product. Maximum likelihood analysis of these data indicate that between 125 and 220 different serine protease genes are expressed in first-instar larvae of L. cuprina. DNA sequence analysis of ten randomly-selected clones, derived from the three tissue sources, indicated that all ten encoded serine protease gene fragments. A frequently occurring PCR product, generated from both first-instar total cDNA and third-instar cardia cDNA, showed 73% amino acid identity to a digestive protease expressed in Droso-phila melanogaster larval gut cells. 相似文献
18.
Gut-specific genes from the black fly Simulium vittatum encoding trypsin-like and carboxypeptidase-like proteins 总被引:3,自引:1,他引:2
In haematophagous insects digestion of the blood meal provides nutrients for survival and essential components for egg production. We have isolated and partially characterized two gut-specific genes from the black fly Simulium vittatum. Sequence analysis revealed that both are highly similar to digestive proteases, one to trypsins and the other to carboxypeptidases. RNA blot analysis indicates that the expression of these two genes is regulated in a sexspecific manner; when fed the same sucrose-based diet, expression in males is substantially lower than in females. In females, expression of both genes is strongly induced by a blood meal. At 6 h after the blood meal the trypsin-like gene product was immunolocalized to the midgut epithelium and to the outer layers of the peritrophic matrix. 相似文献
19.
Cloning and sequence of a cDNA for a highly basic protease from the digestive juice of the silkworm, Bombyx mori 总被引:3,自引:0,他引:3
E. Kotani T. Niwa M. Tokizane K. Suga Y. Sugimura K. Oda H. Mori T. Furusawa 《Insect molecular biology》1999,8(2):299-304
A serine protease of the silkworm, Bombyx mori, with an isoelectric point of pH 10–11 and a pH optimum for succinyl-Leu-Leu-Val-Tyr-MCA degrading activity of about 10, was found in a 0.33 m NaCI-eluted fraction obtained from cation-exchange chromatography of digestive juice. The activity of the enzyme was strongly inhibited by chymostatin and PMSF, indicating that the protease is a chymotrypsin-like serine protease. The N-terminal amino acid sequence of the protease was determined, and a full-length cDNA clone (0.92 kbp) which was isolated from a midgut cDNA library was sequenced. The cDNA encodes a pre-proenzyme of 284 amino acids with a pro-segment of 50 amino acids and mature protein of 234 amino acids. From its primary structure, the predicted molecular mass of the mature protein is 24.5 kDa. A sequence comparison of the Bombyx highly basic protease with other serine proteases revealed that this enzyme is a mammalian-type serine protease with a catalytic triad consisting of His45, Asp92 and Ser186. A large number of Arg residues are encoded by the cDNA which may be responsible for its stability and/or function in the alkaline condition, by remaining charged at high pH. 相似文献
20.
Cloning of cDNA for proteinase 3: a serine protease, antibiotic, and autoantigen from human neutrophils 总被引:10,自引:0,他引:10
下载免费PDF全文
![点击此处可从《The Journal of experimental medicine》网站下载免费的PDF全文](/ch/ext_images/free.gif)
D Campanelli M Melchior Y Fu M Nakata H Shuman C Nathan J E Gabay 《The Journal of experimental medicine》1990,172(6):1709-1715
Closely similar but nonidentical NH2-terminal amino acid sequences have been reported for a protein or proteins in human neutrophils whose bioactivities is/are diverse (as a serine protease, antibiotic, and Wegener's granulomatosis autoantigen) but that share(s) several features: localization in the azurophil granules, a molecular mass of approximately 29 kD, reactivity with diisopropylfluorophosphate, and the ability to degrade elastin. We previously purified one such entity, termed p29b. Using a monospecific antibody, we have cloned from human bone marrow a cDNA encoding the complete p29b protein in its mature form, along with pre- and pro-sequences. The predicted amino acid sequence agrees closely with the NH2-terminal sequence obtained previously from purified p29b, as well as with sequences newly obtained from CNBr fragments. The primary structure is highly homologous to elastase, cathepsin G, T cell granzymes, and other serine proteases, and shares both the catalytic triad and substrate binding pocket of elastase. Hybridization of the full-length cDNA with restriction enzyme digests of human genomic DNA revealed only one fragment. This suggests that the closely related species described previously are the same, and can be subsumed by the term used for the first-described activity, proteinase 3. Proteinase 3 is more abundant in neutrophils than elastase and has a similar proteolytic profile and specific activity. Thus, proteinase 3 may share the role previously attributed to neutrophil elastase in tissue damage, and has the potential to function as an antimicrobial agent. 相似文献