Biomedical subjects

R H Fetterer

Publications and source records attributed to R H Fetterer.

At least 19 recordsLinked to original sources

PubMed2005

Heat shock protein 90 genes of two species of poultry Eimeria: expression and evolutionary analysis.

Heat shock protein 90 (Hsp90) is 1 of the most abundant and evolutionarily conserved proteins. In most species, Hsp90 is essential for proper cell function. In this study, we present the molecular analysis of Hsp90 from Eimeria species, the causative agents of avian coccidiosis. The full-length Eimeria acervulina Hsp90 complementary DNA was isolated from intestinal intraepithelial lymphocytes of Eimeria-infected chickens. From evolutionary analysis and sequence identity, it is likely that Eimeria Hsp90 sequences described thus far encode the cytosolic versions of the protein. Although at the nucleotide and amino acid levels Eimeria tenella and E. acervulina Hsp90 are highly similar, their expression profiles differ considerably. Although E. tenella transcripts were detected in all developmental stages tested, E. acervulina transcripts were not found in oocysts undergoing sporulation or in fully sporulated oocysts, suggesting that messenger RNA expression may be regulated quite differently between Eimeria species.

Amino Acid Sequence↗

PubMed2005

Partial purification and characterization of an aminopeptidase from Eimeria tenella.

Our previous investigation demonstrated the expression in Eimeria tenella sporulated oocysts of an aminopeptidase (AP) with strong homology to AP N. To further understand the role of proteases during development, we investigated the molecular and biochemical properties of E. tenella AP. Greater than 95% AP activity was present in a soluble extract during sporulation of oocysts with highest activity in fully sporulated oocysts. The AP activity was inhibited by the AP inhibitors bestatin and 1,6-phenanthroline, but not by serine protease inhibitors. The AP had specificity for synthetic endopeptidase substrates that contain arginine, alanine, or glycine at the N terminus. Partial purification of the enzyme yielded a major protein band with an Mr of about 106 kDa and an isoelectric point (Ip) of 5.1. Reverse transcription-polymerase chain reaction indicated that the gene for AP is expressed during sporulation, but expression is absent or greatly reduced in the sporozoites and merozoites. On the basis of the deduced gene structure, the predicted Mr is 110 kDa with a pI of 5.59. Database search indicates that the E. tenella AP shares significant homology with the AP from Apicomplexan taxa: Toxoplasma gondii, Cryptosporidium parvum, and Cryptosporidium hominis. Together, these results confirm the presence of a cytosolic AP related to AP N, which is expressed and active during sporulation of E. tenella oocysts.

Amino Acid Sequence↗

PubMed2004

A conserved 19-kDa Eimeria tenella antigen is a profilin-like protein.

A wide range of recombinant proteins from Eimeria species have been reported to offer some degree of protection against infection and disease, but the specific biological function of these proteins is largely unknown. Previous studies have demonstrated a 19-kDa protein of unknown function designated SZ-1 in sporozoites and merozoites of Eimeria acervulina that can be used to confer partial protection against coccidiosis. Reverse transcriptase-polymerase chain reaction indicated that the gene for SZ-1 is expressed by all the asexual stages of Eimeria tenella. Rabbit antisera to recombinant SZ-1 recognized an approximately 19-kDa protein from extracts of E. tenella sporozoites, merozoites, sporulated oocysts, and oocysts in various stages of sporulation. Immunofluorescence antibody staining indicated specific staining of E. tenella sporozoites and merozoites. Staining was most intense in the cytoplasm of the posterior end of the parasite. The primary amino acid sequence of the gene for E. tenella SZ-1 deduced from the E. tenella genome indicated a conserved domain for the actin-regulatory protein profilin. A conserved binding site for poly-L-proline (PLP), characteristic of profilin was also observed. SZ-1 was separated from soluble extract of E. tenella proteins by affinity chromatography using a PLP ligand, confirming the ability of SZ-1 to bind PLP. SZ-1 also partially inhibited the polymerization of actin. The current results are consistent with the classification of SZ-1 as a profilin-related protein.

Actins↗

PubMed2004

Analysis of transcripts expressed by Eimeria tenella oocysts using subtractive hybridization methods.

To characterize the genes expressed by Eimeria tenella oocysts, the sequence of 499 expressed sequence tags (ESTs) was obtained from complementary DNA (cDNAs) enriched for transcripts expressed by unsporulated or sporulated oocysts. Of these, 225 clones were isolated from cDNA of sporulated oocysts and 274 from unsporulated oocysts. A total of 163 unique sequences were found, and the majority of these (64%) represent novel genes with no significant homology to the proteins in GenBank. Approximately half of the unique transcripts generated from sporulated oocysts are also expressed by sporozoites and merozoites, whereas the expression of most (79%) of the transcripts from unsporulated oocysts has not yet been detected at other stages of development. The expression of 4 transcripts obtained from the subtracted cDNAs was confirmed by quantitative reverse transcriptase-polymerase chain reaction. The results confirmed that these transcripts are in fact differentially expressed between sporulated and unsporulated oocysts.

Animals↗

PubMed2003

The effect of dietary betaine on intestinal and plasma levels of betaine in uninfected and coccidia-infected broiler chicks.

Chicks fed betaine supplemented diets and infected with Eimeria acervulina and Eimeria maxima had markedly higher levels of betaine in the duodenum and mid-gut than unsupplemented, infected chicks. Uninfected chicks fed betaine exhibited almost twice the levels of betaine in the gut as infected chicks. Plasma betaine levels were lower in E. maxima-infected chicks than in E. acervulina-or Eimeria tenella-infected chicks. Betaine supplementation reversed the decrease in weight gain in E. maxima- infected chicks but had no effect on the decrease in weight gains in E acervulina- and E. tenella-infected chicks. Coccidia-infected birds on normal diets regularly exhibit increases in plasma NO(2)(+)NO(3). This increase was abolished in E.tenella-infected birds on betaine supplement. Betaine feeding did not alter this effect in E. acervulina- and E. maxima-infected birds. Results indicate that betaine supplementation has a positive effect on gut betaine levels in birds infected with E. acervulina and E. maxima. In all treatment groups, infection lowered the levels of betaine.

Animals↗

PubMed2003

Characterization of a developmentally regulated oocyst protein from Eimeria tenella.

Changes in proteins during sporulation of Eimeria tenella oocysts were investigated. Unsporulated E. tenella oocysts collected from cecal tissue at 7 days postinoculation were sporulated in aerated media at 28 C for 0-48 hr. Gel analysis of soluble protein extracts prepared from oocysts from their respective time points indicated the presence of 2 prominent bands with relative molecular weight (Mr) in the range of 30 kDa and making up 20% of the total protein. These 2 bands, designated as major oocyst proteins (MOPs), were absent or barely detectable by 21 hr of sporulation. MOP bands were weakly reactive with glycoprotein stain but showed no mobility shift on deglycosylation. By gel analysis it was shown that the purified MOPs consisted of 2 bands of Mr 28.7 and 30.1 kDa. However, by matrix-assisted laser deabsorption-time of flight analysis it was shown that masses were about 17% lower. Internal sequence analysis of the 28.7-kDa protein generated 2 peptides of 17 and 14 amino acids in length, consistent with a recently described protein coded by the gam56 gene and expressed in E. maxima gametocytes. Rabbit antibodies made against MOPs were localized to outer portions of sporocysts before excystment and to the apical end of in vitro-derived sporozoites. These same antibodies were found to react with bands of Mr 101 and 65 kDa by Western blot but did not recognize MOPs in soluble or insoluble sporozoite extracts. The data suggest that the MOPs are derived from part of a gametocyte protein similar to that coded by gam56 and are processed during sporulation into sporocyst and sporozoite proteins. Alternatively, the binding of anti-MOP to 101- and 65-kDa proteins may result from alternatively spliced genes as the development of parasite proceeds.

Amino Acid Sequence↗

PubMed2002

Interaction of dietary vitamin E with Eimeria maxima infections in chickens.

In two trials, broiler chickens, processed similarly to those placed in commercial operation, were fed, from 1 d of age, a range (13 to 200 ppm) of DL-alpha-tocopheryl acetate (VE-AC) levels, and the effects on the pathology of Eimeria maxima infections were assessed at 6 d postinoculation (PI). In Trial 1, dietary levels of VE-AC had little significant effect on variables characterizing pathology except for the number of oocysts shed, which was significantly increased in chicks treated with higher VE-AC levels. The infection was judged to be mild based on moderate lesion scores (2.2+/-0.2), lack of significant effects on weight gain (7+/-1.6% decrease), moderate reduction in plasma carotenoids (21+/-2%) and small increases in plasma NO2-+NO3- (141+/-12%). In uninfected and infected chickens, plasma alpha-tocopherol (AT) increased with dietary levels of VE-AC; however, E. maxima infection caused a fairly constant decrease in AT of 35.3+/-3.2% across these levels. Plasma gamma-tocopherol (GT) levels were unaffected by dietary VE-AC or E. maxima infection. In Trial 2, pathology, again, was relatively unaffected by dietary VE-AC level. The infection was judged to be severe based on lesion scores (3.5+/-0.1), reduction in weight gain (30.7+/-3%), plasma carotenoids (72.4+/-1.5%), uric acid (16.3+/-3.4), albumin (37.8+/-2.8%), large increases (261+/-8%) in plasma NO2-+NO3-, and high numbers of oocysts shed per chick (4.12+/-0.4 x 10(7)). Plasma AT again increased with increasing dietary VE-AC levels in uninfected and infected chicks, but the mean decrease across VE-AC levels caused by E. maxima infection was 73.14+/-3.3%. GT levels were erratic and unrelated to dietary VEAC or infection. Thus, in processed broiler chickens, high dietary VE-AC did not prevent or lessen the pathology caused by mild or severe infections with E. maxima. The main effect of E. maxima infection appeared to be reduction in plasma AT levels. We postulate that this reduction may be due to malabsorption of AT, which results from physical damage to the absorptive mucosa, reduction in esterases required to hydrolyze the VE-AC, and a generalized lipid malabsorption, preventing movement of the free AT to circulating blood and infected tissues.

Animal Feed↗

PubMed2002

Recent advances in biology and immunobiology of Eimeria species and in diagnosis and control of infection with these coccidian parasites of poultry.

Avian coccidiosis, an intestinal disease caused by protozoan parasites of the genus Eimeria, occurs worldwide. It is considered to be one of the most economically important diseases of domestic poultry. For many years, prophylactic use of anticoccidial feed additives has been the primary means of controlling coccidiosis in the broiler industry and has played a major role in the growth of this industry, which now can produce about 7.6 billion chickens annually. However, development of anticoccidial resistance has threatened the economic stability of the broiler industry. Although there has been little effort by the pharmaceutical industry to develop new anticoccidials, the mounting problem of drug resistance of Eimeria species has prompted major research efforts to seek alternative means of control through increased knowledge of parasite biology, host response, and nutritional modulation. As a consequence, important advancements have been made, particularly in defining parasite antigens that have potential use in vaccines, defining the Eimeria genome, understanding the immunology of coccidial infections, and the practical applications of live vaccines. This review describes the progress in these areas, most of which has occurred within the past 10 to 15 years.

Animals↗

PubMed2001

Release of hyaluronidase during in vitro development of Ascaris suum from the third to fourth larval stage.

An enzyme that degraded glycosaminoglycan hyaluronic acid was released during in vitro development of Ascaris suum L3 to L4. The enzyme did not hydrolyze glycosaminoglycan chondroitin sulfate A. One molecular form of hyaluronidase was detected, with a molecular weight estimated at 47.8 +/- 8.6 kDa by sucrose density gradient centrifugation and at 55.0 +/- 1.3 kDa by substrate SDS-PAGE zymography. Activity of the enzyme was optimal between pH 5.0 and 6.0, and was present at neutral pH. Hyaluronidase activity was not affected by 5 mM concentrations of cupric sulfate, zinc chloride, calcium chloride, manganese chloride or EDTA. In addition, NaCl had no effect on enzyme activity at concentrations of 0.2-1.0 M. The highest level of hyaluronidase was present in culture fluid collected between days 4 and 6 of in vitro culture, and this period corresponded with that of the highest rate of increase in the percentage of L4. The presence or absence of hyaluronic acid plays a key role in basic developmental processes of vertebrates and is regulated, in part, by hyaluronidases. Developmental processes occurring during the transition of A. suum L3 to L4 may likewise depend on hyaluronidase. In addition, the infection process of a number of organisms, including some nematodes, depends on hyaluronidase. A. suum may likewise utilize hyaluronidase to facilitate larval migration within the host.

Animals↗

PubMed2001

Eimeria tenella infection in chickens: effect on plasma and muscle 3-methylhistidine.

To assess muscle breakdown during avian coccidiosis, the level of the nonmetabolizable amino acid 3-methylhistidine (3MH) was determined in muscle and plasma from chickens infected with the cecal parasite Eimeria tenella. The change in 3MH level during infection was determined in birds, each inoculated with 0 to 200,000 sporulated oocysts. The effect of levels of parasitism was evaluated at 6 d postinoculation. The 3MH levels of plasma and muscle were determined by HPLC after derivatization with fluorescamine. Weight gains, packed cell volumes, and gross lesion scores were also determined. E. tenella infected birds with lesion scores of 3 or 4 had significantly elevated plasma and muscle 3MH, whereas infected birds with lesion scores of 0, 1, or 2 did not have elevated plasma and muscle 3MH; however, there was a linear inverse relationship between weight gain and both plasma and muscle 3MH. The results suggested that muscle breakdown, as assessed by plasma and muscle levels of 3MH, was elevated during the acute stage of E. tenella infection and was most likely associated with anorexia caused by infection. However, the correlation of 3MH levels with severity of infection was not as strong as that previously observed for E. acervulina infection, most likely due to the differences in pathology caused by the two species.

Animals↗

PubMed2001

Cuticular collagen synthesis by Ascaris suum during development from the third to fourth larval stage: identification of a potential chemotherapeutic agent with a novel mechanism of action.

The dominant proteins released by Ascaris suum during development in vitro from the L3 to L4 stage were identified as collagenous cuticular proteins by sequence analysis and susceptibility to digestion by collagenase. Under reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the collagen proteins separated into 3 groups with molecular weights estimated at 32 kDa, 54-60 kDa, and 71-91 kDa. The 32-kDa protein represents monomeric collagen; the 54-60- and 71-91-kDa components represent dimeric and trimeric forms, respectively, polymerized by nonreducible cross-links. Furthermore, the release of these forms of collagen was developmentally regulated, as exemplified by a sequential temporal progression from monomeric to dimeric to trimeric forms in association with the in vitro transition from L3 to L4. The data suggest that collagen released in vitro during development of A. suum L3 to L4 reflects the increased translation of collagen gene products and their initial assembly into higher molecular weight molecules associated with the synthesis of the L4 cuticle. A biotinylated dipeptidyl fluoromethylketone cysteine protease inhibitor (Bio-phe-ala-FMK) bound specifically to the 32-kDa collagen and, to a lesser extent, to a 30-kDa protein; binding was dependent on the presence of dithiothreitol (DTT) and was prevented by iodoacetamide. Because cysteine residues play an essential role in the initial assembly of the collagen monomers into the higher molecular weight oligomers present in the mature nematode cuticle, inhibition of molting of A. suum L3 to L4 by the cysteine protease inhibitor Z-phe-ala-FMK might be due to its binding to thiol groups of collagen monomers during a critical phase of collagen assembly. Prevention of cystine cross-links during this critical period of cuticle assembly by peptide-FMK inhibitors may represent a potential control mechanism having a novel mechanism of action.

Amino Acid Sequence↗

PubMed2000

Trichuris suis: A secretory serine protease inhibitor.

A trypsin inhibitor was identified in extracts of adult Trichuris suis and culture fluids from 24-h in vitro cultivation of adult parasites. The inhibitor was isolated by acid precipitation, affinity chromatography (trypsin-agarose), and reverse phase HPLC as a single polypeptide with a molecular weight estimated at 6.6 kDa by laser desorption mass spectrometry. The purified inhibitor associated strongly with trypsin (equilibrium dissociation inhibitory constant (K(j)) of 3.07 nM) and chymotrypsin (K(j) = 24.5 nM) and was termed TsTCI. Elastase, thrombin, and factor Xa were not inhibited. The cDNA-derived amino acid sequence of the mature TsTCI consisted of 61 residues including 8 cysteine residues with a molecular mass of 6.687 kDa. The N-terminal region of TsTCI (46 residues) showed limited homology (36%) to a protease inhibitor from the hemolymph of the honeybee Apis mellifera, which is considered to be a member of the Ascaris inhibitor family. However, TsTCI did not display sequence homology with other members of this family or the distinctive cysteine residue pattern which distinguishes this family. However, similarity of a region of TsTCI (11 residues) with the reactive site regions of inhibitors from the nematodes Ascaris suum, Anisakis simplex, and Ancylostoma caninum was apparent.

Amino Acid Sequence↗

PubMed2000

Trichuris suis: a secretory chymotrypsin/elastase inhibitor with potential as an immunomodulator.

A serine protease inhibitor, termed TsCEI, was purified from adult-stage Trichuris suis by acid precipitation, affinity chromatography (elastase-agarose), and reverse-phase HPLC. The molecular weight of TsCEI was estimated at 6.437 kDa by laser desorption mass spectrometry. TsCEI potently inhibited both chymotrypsin (K(i) = 33.4 pM) and pancreatic elastase (K(i) = 8.32 nM). Neutrophil elastase, chymase (mouse mast cell protease-1, mMCP-1), and cathepsin G were also inhibited by TsCEI, whereas trypsin, thrombin, and factor Xa were not. The cDNA-derived amino acid sequence of the mature TsCEI consisted of 58 residues including 9 cysteine residues with a molecular mass of 6.196 kDa. TsCEI displayed 48% sequence identity to a previously characterized trypsin/chymotrypsin inhibitor of T. suis, TsTCI. TsCEI showed 36% sequence identity to a protease inhibitor from the hemolymph of the honeybee Apis mellifera. Sequence similarity was also detected with the trypsin/thrombin inhibitor of the European frog Bombina bombina, the elastase isoinhibitors of the nematode Anisakis simplex, and the chymotrypsin/elastase and trypsin inhibitors of the nematode Ascaris suum. The inhibitors of T. suis, an intestinal parasite of swine, may function as components of a parasite defense mechanism by modulating intestinal mucosal mast cell-associated, protease-mediated, host immune responses.

Amino Acid Sequence↗

PubMed2000

Effect of Eimeria acervulina infections on plasma L-arginine.

As part of a program to study the pathological effects of coccidia infections on growth, we have examined the relationship of plasma L-arginine (ARG) levels to infective doses of Eimeria acervulina and infection-associated changes in weight gain, plasma carotenoids, and plasma NO2- + NO3-. Chickens consuming a starter ration containing 1.68% ARG were infected with a range of doses of E. acervulina. At 6 d postinoculation (PI), weight gains were significantly reduced by infections with 5 x 10(5) and 1 x 10(6) oocysts per chick (OPC). Gross lesion scores of chickens infected with 5 x 10(4) through 1 x 10(6) OPC were significantly greater than scores of chicks infected with 1 x 10(3) OPC. Compared with levels from uninfected controls, plasma NO2- + NO3- concentrations were significantly increased by infection with 5 x 10(5) and 1 x 10(6) OPC, plasma concentrations of ARG were significantly decreased by infection with 5 x 10(4) through 1 x 10(6) OPC, and plasma carotenoids were significantly decreased by all infection doses. Plasma arginine was significantly correlated with plasma carotenoids (P > 0.0187), but not with infection dose or weight gain; plasma NO2- + NO3- was positively correlated (P > 0.0043) with infection dose and negatively correlated (P > 0.0158) with weight gain. Regression analysis of the measured variables indicated that the strongest relationship existed between plasma ARG and carotenoids. This finding suggests that in this infection model, reduction in plasma ARG is most likely associated with nutrient malabsorption that accompanies infection and is likely not significantly impacted by synthesis of nitric oxide that is associated with the immune response.

Animals↗

PubMed2000

Characterization of acid phosphatase and phosphorylcholine hydrolase in adult Haemonchus contortus.

An acid phosphatase (AP) and a phosphorylcholine hydrolase (PCH) were detected in excretory-secretory (ESP) products from adult Haemonchus contortus. The AP had a pH optimum of 4.5 and was inhibited by tartaric acid and sodium fluoride, but not by o-phenanthroline. The AP hydrolyzed paranitrophenol (pnp)-phosphate and to a lesser extent pnp-phenyl-phosphonate but did not hydrolyze diester substrates. Purified AP consisted of heterodimers with relative molecular weight (Mr) of 41.9 and 48.7 kDa and had a native molecular weight of 98 kDa by size-exclusion chromatography (SEC). The PCH had a pH optimum of about 9.5 and was inhibited by EDTA and o-phenanthroline but not by the specific phospholipase inhibitor D609. The specific activity of PCH in the ESP was approximately 25-fold less than that of AP. PCH also hydrolyzed 5'-thymidine monophosphate-pnp at a rate about 40% lower than pnp-phosphorylcholine but did not hydrolyze 3'-thymidine monophosphate-pnp. Partial purification of PCH suggests an Mr of 50.2 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and an Mr of 102 kDa by SEC. Both AP and PHC were secreted in vitro in a time-dependent manner and had their highest concentrations in the intestine. The results indicate that H. contortus adults secrete significant amounts of AP that might be a digestive enzyme. PCH is also an intestinal enzyme and is secreted in lesser amounts than AP. The PCH is probably not a phospholipase C but has some characteristics of a type I phosphodiesterase.

Acid Phosphatase↗

PubMed2000

Eimeria acervulina infection elevates plasma and muscle 3-methylhistidine levels in chickens.

To assess muscle breakdown during avian coccidiosis, the level of the nonmetabolizable amino acid 3-methylhistidine (3MH) was determined in muscle, plasma and excreta from chickens infected with Eimeria acervulina. The changes in 3MH levels during infection were assessed at 1-29 days postinoculation (DPI) in animals given 5 x 10(5) oocysts per bird. The effect of levels of parasitism were evaluated at 8 DPI in birds receiving 5 x 10(3), 5 x 10(4), 5 x 10(5) or 1 x 10(6) oocysts each. The 3MH levels of plasma, muscle, and excreta samples were determined by high-pressure liquid chromatography after derivatization with fluorescamine. Weight gains, breast muscle weight, eviscerated weight, plasma carotenoid levels, dry weight of muscle, and gross lesion scores were also determined. Infected birds had significantly elevated plasma and muscle 3MH at 4 and 8 DPI following a single dose of E. acervulina. The increase in 3MH levels had an inverse relationship with the time course of weight gain and plasma carotenoid levels. Plasma and muscle 3MH levels returned to control values by 15 DPI and remained unchanged from control values through the remainder of the experiment (29 DPI). Breast weight was decreased in infected birds, but the ratio of breast weight to eviscerated body weight was unchanged. Excretion of 3MH decreased relative to controls at 4 and 8 DPI and returned to control levels on 15 DPI. The plasma and muscle levels of 3MH were related to severity of infection; however, levels of excreted 3MH were not. The results suggested that muscle breakdown, as assessed by plasma and muscle levels of 3MH, increased during the acute stage of E. acervulina infection. The underlying causes for this muscle breakdown was unclear but could involve a physiological response to anorexia and decreased food intake during the acute phase of infection. Levels of excreted 3MH did not increase during infection and this may be the result of decreased excreta output during infection. Plasma and muscle levels of 3MH were correlated with severity of E. acervulina infections but may not be as sensitive an indicator of infection as plasma carotenoid levels or other physiological parameters.

Animals↗

PubMed2000

A developmentally regulated hyaluronidase of Haemonchus contortus.

The trichostrongylid nematode Haemonchus contortus released a hyaluronic acid-degrading enzyme during in vitro development from the third (L3) to fourth (L4) larval stage. The enzyme did not degrade chondroitin sulfate A. Enzyme activity was optimal between pH 4.0 and 6.0, and the enzyme was inhibited by high concentrations of NaCl; the divalent cations Cu2+, Zn2+, Ca2+, and Mn2+ were not inhibitory. The hyaluronidase had a molecular mass estimated at 57 kDa by sucrose density gradient centrifugation and at 111 kDa by substrate sodium dodecyl sulfate polyacrylamide gel electrophoresis (reducing and nonreducing conditions), suggesting the formation of a dimer during the electrophoretic separation conditions. The level of hyaluronidase released during in vitro development peaked between 24 and 48 hr in culture and then gradually decreased, with little or no activity present in the 168-hr culture fluid. The enzyme was not detected in culture fluid from 24-hr incubations of either the mid-L4 stage (obtained from sheep 7 days postinfection) or the adult stage (obtained from sheep 30-35 days postinfection). The temporal expression of the hyaluronidase suggested a role for this enzyme in the early stages of the L3-L4 developmental process.

Animals↗

PubMed1999

Characterization of a hemoglobin-like protein from adult Haemonchus contortus.

A hemoglobin-like protein was purified from supernatants of adult Haemonchus contortus extracts by high-pressure liquid chromatography. The purified protein had an M(r) of 33 kDa as determined by size-exclusion chromatography under non-denaturing conditions and an M(r) of 19 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting the hemoglobin may exist as a dimer. The sequences of 3 peptides resulting from proteolytic digest of the purified protein were determined and demonstrated greater than 50% identity to the globin from Trichostrongylus colubriformis. Adult H. contortus incubated overnight in [3H]leucine, incorporated radioactivity into a peak that coeluted with parasite hemoglobin, indicating the adults synthesize hemoglobin in vitro. The L3-stage lacked hemoglobin, but the L4-stage contained a hemoglobin with an M(r) of 19.6 kDa.

Amino Acid Sequence↗