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Biomedical subjects

C L Ho

Publications and source records attributed to C L Ho.

At least 73 records · Page 4Linked to original sources
PubMed1995

Immunogenicity of mastoparan B, a cationic tetradecapeptide isolated from the hornet (Vespa basalis) venom, and its structural requirements.

Mastoparan B (MP-B) is a cationic tetradecapeptide (LKLKSIVSWAKKVL-CONH2, mol. wt 1611) isolated from the black-bellied hornet (Vespa basalis) venom. The small peptide itself was capable of inducing antibodies without prior conjugation to a protein carrier in rabbits and mice. The mouse antibody was found to be of IgG1 isotype with kappa-type light chain. The peptide antigen was able to form insoluble complexes with the specific antibody, suggesting that MP-B possessed more than one epitope in its molecule. The finding that MP-B was able to bind with both mouse and rabbit antibodies in sandwich ELISA supports this contention. Synthetic MP-B analogues in which lysine at position 2, 4, 11 or 12 was replaced by neutral amino acids such as asparagine or leucine showed a significant decrease in their antibody-binding activities. Substitution of lysine at position 4 (Lys4) caused the most marked inhibition in its binding activity. However, replacing tryptophan at position 9 by tyrosine caused a relatively small reduction in its binding activity. Replacing both Lys2,4 by asparagine or removing Lys-containing segments at amino or carboxyl terminus in MP-B sequence caused a remarkable decrease in the antibody-binding and immunogenic activities of the peptide. The Lys residues located at amino and carboxyl terminal segments of MP-B, especially Lys4, appear to play a critical role in the binding interaction and the immunogenicity of the peptide.

Amino Acid Sequence↗
PubMed1995

Inhibition of protein kinase C by snake venom toxins: comparison of enzyme inhibition, lethality and hemolysis among different cardiotoxin isoforms.

Cardiotoxins, neurotoxins and phospholipase A2 (PLA2) are three major classes of toxic components present in the Taiwan cobra, Naja naja atra of the Elapidae family. Cardiotoxins (or called cytotoxins), a group of major polypeptides of around 60 amino-acid residues present abundantly in the elapid family of snakes, comprise about 45-55% of the crude venom of Taiwan cobra. In contrast to another prominent group of structurally similar neurotoxins with well-established acetylcholine receptors and modes of action, cardiotoxins showed no defined cellular targets and very diverse pharmacological functions. A systematic structure/function comparison of these toxins was made by their relative inhibitory effects on protein kinase C (PKC) isolated from mouse brains. Lethality and hemolysis of various cardiotoxin isoforms were also compared in order to shed some insight on the biological targets and mechanisms of these surface-active amphiphilic polypeptides. A structure comparison of these cardiotoxins based on computer model-building revealed that some defined and subtle differences can be detected upon the superposition of these three-dimensional polypeptide chains, which may reflect the intrinsic differences in the hydrophobic peptide segments present on the surface loops of toxin molecules. The differences seem to correlate with different inhibitory activities exhibited by cardiotoxins in contrast to the lack of activity by cobrotoxin and PLA2 on PKC.

Animals↗
PubMed1995

Translation initiation and assembly of peripherin in cultured cells.

The peripherin gene has three potential ATG translation initiation sites at positions 38, 56, and 290. The second ATG has been proposed to be the initiation codon used for translation of the protein, but there is no experimental evidence for this conjecture. We have isolated a full-length peripherin cDNA (designated as p61-11) from a rat brain cDNA library. Upon sequencing, we found that this cDNA contains a point mutation at the second potential translation initiation codon, which changes this ATG to ACG. When expressed in SW13 cl.2 vim- cells, a cell line without any detectable cytoplasmic intermediate filaments, the protein product of p61-11 cannot form a filamentous network and the major product is 45 kDa in size, which is most likely initiated from the third ATG. The protein product from the first ATG (57 kDa in size) of p61-11 is also detected albeit in smaller amounts. We introduced a frame-shift mutation upstream of the third ATG in p61-11 to create p61-11FS and showed that the third ATG is able to initiate translation efficiently even in the presence of the first ATG, and the 45 kDa protein leads to a diffuse nonfilamentous staining pattern in vim- cells confirming that the first ATG may not be the preferred translation initiation codon, since it cannot suppress a downstream ATG. We increased the translation efficiency from the first ATG of p61-11 by mutating the three nucleotides preceding this first ATG and thereby placing it in a better Kozak consensus sequence for translation initiation. The resulting 57 kDa protein is able to form a filamentous network in vim- cells. We corrected the mutation in the original p61-11 by polymerase chain reaction and generated two peripherin constructs: perM1M2 (which contains all three translation initiation codons) and per delta 1M2 (the first ATG is deleted, but the other two are present). When transfected, their protein products, about 57 kDa in size, form filamentous networks in the absence of other cytoplasmic intermediate filaments. Since there is no 45 kDa protein detected for these latter two constructs, it is reasonable to conclude that in the presence of the second ATG, little or no translation is initiated from the third ATG. Taken together, these results strongly suggest that the second ATG is the preferred translation initiation codon for the peripherin gene.

Adenocarcinoma↗
PubMed1994

Cardiovascular effects of mastoparan B and its structural requirements.

Mastoparan B is a cationic, amphiphilic tetradecaeptide (LKLKSIVSWAKKVL-CONH2) isolated from the venom of the hornet Vespa basalis. Intravenous injection of the peptide into rats caused a profound depression of blood pressure and cardiac function, which was inhibited by cyproheptadine, reserpine and multiple doses of compound 48/80, but not by diphenhydramine and cromolyn. Mastoparan from Paravespula lewisii showed little cardiovascular inhibitory activity. A synthetic mastoparan B analog in which lysine at position 2 was replaced by asparagine showed a marked decrease in the cardiovascular depressor effects, while replacing lysine at position 4, 11 or 12 with leucine did not cause a significant reduction in these effects. Replacing lysine at position 12 with leucine even caused a more sustained depressor effect. However, the analog in which lysines at positions 11 and 12 were replaced by leucine lost its cardiovascular inhibitory activity. Replacing tryptophan at position 9 with phenylalanine in mastoparan B did not affect its activity. It is concluded that mastoparan B is involved in the cardiovascular disturbances induced by the hornet venom. Lysine at position 2 is a critical residue for the cardiovascular effects of mastoparan B. A peptide molecule with two lysine residues, one located close to the amino terminus and the other near the carboxyl end of the peptide, appears to be the optimal structure for eliciting the cardiovascular depressor effects.

Amino Acid Sequence↗
PubMed1994

Vitreous fluorophotometry in patients with crystalline retinopathy.

Patients with various stages of crystalline retinopathy were examined with vitreous fluorophotometry and the results were compared with those of normal subjects. Two patients with glistening crystals without pigment clumps, together with atrophy of the retinal pigment epithelium in the posterior pole, showed a normal inward permeability of fluorescein. Three patients with a large area of choriocapillaris atrophy in the posterior pole, in addition to atrophy of retinal pigment epithelium, demonstrated an increase in the rate of inward fluorescein penetration. Three patients with extensive areas of choriocapillaris atrophy throughout the fundus also revealed an increased posterior vitreous penetration ratio.

Adult↗
PubMed1993

Structural requirements of mastoparan for activation of membrane-bound guanylate cyclase.

Mastoparan activated membrane-bound guanylate cyclase and potentiated the effect of atrial natriuretic factor (ANF) and ATP on guanylate cyclase activity in rat lung membranes. Mastoparan is a cationic, amphiphilic tetradecapeptide with an amidated carboxyl terminus. It takes the alpha-helical conformation upon interacting with the membrane. Several analogs were synthesized to study the role of the positive charges, the carboxyl amino group and the alpha-helical conformation of mastoparan in the activation of guanylate cyclase. The results showed that substitution of the C-terminal amide group of mastoparan with a carboxyl group significantly reduced its potency on the activation of guanylate cyclase. Replacement of three lysine residues of mastoparan with aspartic acid or serine residues completely abolished the stimulatory effect of mastoparan. When the alanine at position 10 of mastoparan was substituted by a proline, the resulting analog had no effect on guanylate cyclase activity. These results demonstrate that the positive charges and the helical structure of mastoparan are critical determinants for the activation of guanylate cyclase.

Amino Acid Sequence↗
PubMed1993

Edema-inducing activity of a lethal protein with phospholipase A1 activity isolated from the black-bellied hornet (Vespa basalis) venom.

The lethal protein of the hornet (Vespa basalis) venom is a phospholipase A1 toxin (mol. wt approximately 32,000) with a potent hemolytic activity. Subplantar injection of the toxin caused a dose-dependent swelling in the rat hind paw. Its potency was higher than those of phospholipases A2 and cardiotoxin from cobra venoms. Hind-paw edema induced by the toxin was inhibited by antiserotonin drugs (cyproheptadine and methysergide), indomethacin and betamethasone. Antihistamine (chlorpheniramine) showed a relatively weak inhibition. Intradermal injection of the toxin into back skin of the rat also induced local edema which was inhibited by chlorpheniramine and methysergide. Rats pretreated with multiple doses of compound 48/80 showed a moderate decrease in the histamine and serotonin content of rat skin, and a slight decrease in paw edema induced by the toxin, while a single dose of reserpine markedly diminished the toxin-induced edema in association with depletion of serotonin in rat skin. The edema-inhibitory action of amine-depleting agents appeared to correlate with their potencies to deplete serotonin in the skin. It is suggested that serotonin, prostaglandin E2, and to a lesser extent of histamine are involved in producing the local effect of the toxin. However, serotonin released by the toxin appears to be the major factor mediating the toxin-induced edema in the rat.

Animals↗
PubMed1993

Mastoparan B, synthesis and its physical and biological properties.

Mastoparan B, a tetradecapeptide toxin (LKLKSIVSWAKKVL) isolated from the hornet (Vespa basalis) venom, was synthesized chemically. The physical and biological properties of both the native and synthetic peptides were studied and proved to be identical. Mastoparan B was found to have a potent antibacterial activity to both Gram negative and Gram positive bacteria.

Amino Acid Sequence↗
PubMed1992

Characterization and molecular cloning of neurotoxic phospholipases A2 from Taiwan viper (Vipera russelli formosensis).

Two phospholipases A2 (PLA2s), designated as RV-4 and RV-7 were purified from venom of the Taiwan Russell's viper (Vipera russelli formosensis) by gel-filtration and reverse-phase HPLC. Their primary structures were solved by both protein sequencing and cDNA cloning and sequencing. The cDNA synthesized was amplified by the polymerase-chain reaction using a pair of synthetic oligonucleotide primers corresponding to the N- and the C-terminal flanking regions of the enzymes. The deduced amino acid sequences of RV-4 and RV-7 were 92% identical to those of the vipoxin and vipoxin inhibitor, respectively, from the Bulgarian Vipera a. ammodytes. RV-4 itself was neurotoxic, whereas RV-7 had much lower enzymatic activity and was not toxic. The low enzymatic activity of RV-7 may be attributed to five acidic residues at positions 7, 17, 59, 114 and 119, which presumably impair its binding to aggregated lipid substrates. Based on the sequence comparison among all the known group II PLA2s, residues 6, 12, 76-81, and 119-125 were identified as important for the neurotoxicity. RV-4 and RV-7 exist in the crude venom as heterodimers, which were again formed by mixing together the HPLC-purified RV-4 and RV-7. Moreover, RV-7 inhibited the enzymatic activity of RV-4 in vitro but potentiated its lethal potency and neurotoxicity. It is suggested that RV-7 may facilitate the specific binding of RV-4 to its presynaptic binding sites, probably by preventing its non-specific adsorption.

Amino Acid Sequence↗
PubMed1991

Structure and biological activities of a new mastoparan isolated from the venom of the hornet Vespa basalis.

By gel filtration on a Fractogel TSK HW 50 column followed by cation-exchange chromatography on CM-Trisacryl M, a tetradecapeptide amide, designated 'mastoparan B', was purified from the venom of the hornet Vespa basalis. Its amino acid sequence was determined as: Leu-Lys-Leu-Lys-Ser-Ile-Val-Ser-Trp-Ala-Lys-Lys-Val-Leu-NH2 and its molecular mass was measured to be 1611 Da by fast-atom-bombardment mass spectrometry. In addition to having a common structure of vespid mastoparans, the peptide shows a less hydrophobic sequence at positions 1, 2, 5, 8 and 9. The peptide caused liberation of histamine from rat peritoneal mast cells and induced oedema in the rat paw. However, the latter effect was inhibited by 'anti-serotonin' (anti-5-hydroxytryptamine) (cyproheptadine), but not by antihistamine (chlorpheniramine). The peptide also possesses a potent haemolytic activity which acts in synergy with the lethal protein of the venom, suggesting the possible involvement of mastoparan B in the lethal effect of Vespa basalis venom.

Amino Acid Sequence↗
PubMed1991

Local edema induced by the black-bellied hornet (Vespa basalis) venom and its components.

The black-bellied hornet, Vespa basalis, is one of the most dangerous species of wasp in Taiwan. The hornet venom possesses a potent edema-inducing activity in addition to its lethal cardiovascular effect. Rat hind-paw edema induced by the venom was inhibited significantly by antiserotonin compounds (cyproheptadine and methysergide) but not by antihistaminics (chlorpheniramine and diphenhydramine), betamethasone, cromolyn, indomethacin plus nordihydroguaiaretic acid, cellulose sulfate, aprotinin or captopril. However, chlorpheniramine or betamethasone, when given in combination with cyproheptadine, showed a significant further inhibition of the venom-induced edema. Pretreating the rat with compound 48/80 reduced the edematous response to the venom. Four venom components, i.e. the lethal protein, mastoparan B, fraction 3 (containing protease) and fraction 15 (containing serotonin) were involved in the edematous effect of the venom. Paw edema induced by the crude venom and its protein components (lethal protein, mastoparan B and fraction 3) was similar in their responses to antiserotonin compounds and antihistamine (inhibited by cyproheptadine and methysergide but not by chlorpheniramine), while that induced by fraction 15 was inhibited by both. It is postulated that upon hornet stings, fraction 15 is acting as an exogenous serotonin, while the protein components, especially the lethal protein, are responsible for the release of endogenous autacoids in which serotonin plays the most important role in inducing local edema.

Animals↗
PubMed1990

Electrophysiological effects of dermorphin on locus coeruleus neurons of rat.

Intracellular recording was used to study the effects of dermorphin on neurons of the locus coeruleus in the rat, in a totally submerged brain slice preparation. Dermorphin caused the inhibition of spontaneous firing of all neurons of the locus coeruleus tested, with an IC50 of 7 nM. Based on the inhibition of spontaneous firing rate, dermorphin was 16.5 times more potent than morphine. Larger concentrations of dermorphin (30-100 nM) further hyperpolarized the neurons of the locus coeruleus and simultaneously caused a reduction in input resistance. These effects were antagonized by naloxone, with a dissociation equilibrium constant of 0.8 nM. The hyperpolarization of neurons of the locus coeruleus, caused by dermorphin, was reversed at a membrane potential of -112 mV in this preparation. Furthermore, this hyperpolarization was blocked by cesium chloride and barium chloride. Thus, these data suggest that dermorphin binds to mu-opioid receptors on the cell membrane of neurons of the locus coeruleus. This leads to opening of the inward-going rectification potassium channels, resulting in the observed hyperpolarization of the membrane.

Analgesics, Opioid↗
PubMed1990

Analgesic, receptor binding, and peripheral opioid activities of synthetic dermorphin-dynorphin hybrid peptides.

The effects of substituting the enkephalin moiety of dynorphin with the dermorphin sequence were studied on the receptor preference, analgesic, and peripheral opioid potencies by using synthetic dermorphin-dynorphin hybrid peptides as the probe. Replacement of the enkephalin moiety of dynorphin with the dermorphin or dermorphin1-5 sequences caused a remarkable increase in analgesic potency, and a 3-6 fold increase in potency of binding against [3H]-dihydromorphine. The potency of receptor binding against [3H]-EKC was also increased by incorporation of the whole dermorphin sequence into the dynorphin molecule. In the presence of NaCl (100 mM), the effect of enhancing binding against [3H]-EKC due to dermorphin substitution disappeared, suggesting the contribution of opioid mu-receptor. Peripheral opioid activities assayed by various smooth muscle preparations showed that dermorphin incorporation caused a decreased in the potency of inhibition of the contractions of the guinea pig ileum and the rabbit vas deferens, no change in potency on the mouse vas deferens, and a marked increase in the inhibition of the rat vas deferens. Among the peripheral opioid activities only that assayed with the rat vas deferens appears to correlate approximately with the analgesic and the receptor binding activities. Judging from the relative potencies obtained from all assays, it is evident that the N-terminal dermorphin moiety, but not the C-terminal dynorphin fragment, dominates the opioid activity and receptor preference of the hybrid peptide.

Amino Acid Sequence↗
PubMed1988

Purification and characterization of a lethal protein with phospholipase A1 activity from the hornet (Vespa basalis) venom.

The hornet, Vespa basalis, is one of the most dangerous species of wasps found in Taiwan. The insect is aggressive and its venom is highly toxic. By gel filtration on a Fractogel TSK HW 50 column followed by cation-exchange chromatography on CM-Trisacryl M, a lethal protein was purified from the venom. It has a molecular mass of about 32 kDa and an i.v. LD50 value of 0.32 micrograms/g mouse. The toxin is capable of catalyzing the hydrolysis of emulsified phospholipids but not sphingomyelin. Analysis of the 1H-NMR spectra of the substrates and its hydrolytic products revealed that the toxin liberates fatty acid from the 1-position of sn-3-phosphoacylglycerols. This result indicates that the toxin possesses phospholipase A1 activity. The toxin exhibits an extremely potent hemolytic activity in washed red cells and diluted whole blood (HC50 = 0.09 micrograms/ml in mouse). The potency of direct hemolysis is about 100-times that of a basic phospholipase A2 from Naja nigricollis venom and about 1000-times that of a cardiotoxin from Naja naja atra venom. A positive correlation between the hemolytic activity and lethality of the toxin was found in three species of animals (mouse, rat and guinea pig). In the in vivo study, the toxin caused a marked increase in the plasma K+ concentration and a hyperkalemic change in the ECG of the treated rat. Hyperkalemia resulting from the hemolytic action of the toxin appears to be the main cause of death in the animal.

Amino Acid Sequence↗
PubMed1988

Beta-endorphin. Receptor binding and peripheral opioid activities of [Gln8]-, [Trp27]-, and [Tyr31]-analogs.

Three beta h-EP analogs which show different extents of alteration in analgesic potency by substitution of a single amino acid residue were assayed for their peripheral opioid activity and the binding to opioid mu-receptor to determine the relationships among the opioid activities obtained from different assays. In the guinea pig ileum assay, [Gln8]-beta h-EP showed a higher inhibitory activity than the parent peptide, [Tyr31]-analog had the same potency as beta h-EP, while [Trp27]-analog retained only one fourth the potency of beta h-EP. Assayed on the vas deferens of the mouse and the rat, all three substituted beta h-EP analogs exhibited a lower potency than their parent peptide. Receptor binding assay using [3H]-dihydromorphine as the primary ligand showed that [Gln8]-analog had a binding potency 1.5-fold that of beta h-EP, while the potencies of [Tyr31]- and [Trp27]-analogs were not significantly different from that of the parent peptide. No correlation in relative potency was found between vas deferens assays and their mu-receptor binding or analgesic activity. However, the relative potencies of binding to mu-receptor in [Gln8]- and [Tyr31]-analogs were found to be consistent with those of analgesic and guinea pig ileum assays, whereas the binding to beta-EP receptor of all analogs appeared to be related to the charge properties of beta-EP molecule.

Animals↗
PubMed1987

Effects of equinatoxin on the guinea-pig atrium.

Equinatoxin is a lethal protein isolated from a sea anemone, Actinia equina. The toxin (0.1-3 micrograms/ml) caused an initial inhibition followed by an augmentation of contractions and beating rates in the isolated guinea-pig atrium. The inhibitory phase was transient (30-60 sec), while the stimulant phase lasted for about 30 min. The treated atrium showed tachyphylaxis to the toxin. The inhibitory effect of the toxin was diminished by tetrodotoxin and atropine and abolished by 8-phenyltheophylline or mepacrine. Dipyridamole, which blocks the uptake of adenosine in the heart, enhanced the inhibitory effect. The stimulant effect of the toxin was inhibited by indomethacin or mepacrine and abolished by a combination of both, but was not inhibited by propranolol. Bioluminescent assay performed during the inhibitory phase showed an increased release of ATP and radioimmunoassay during the stimulant phase revealed an increased release of prostaglandin E2 from the treated atrium. These results suggest that the cardiac inhibitory effect of equinatoxin is mainly due to release of adenyl compounds, while the cardiac stimulant effect of the toxin may result from the liberation of arachidonic acid and subsequent formation of prostaglandins in the guinea-pig atrium.

Adenosine Triphosphate↗
PubMed1987

Beta-endorphin. Biological activity of analogs containing dermorphin and dynorphin sequences: ileum and vas deferens assays.

Biological activity of synthetic beta-endorphin (beta-EP) analogs containing dermorphin or dynorphin-A-(1-13) structure has been investigated using the guinea pig ileum and the vas deferens of the mouse, rat and rabbit. Replacement of NH2-terminal 1-7 segment of camel beta-EP [beta c-EP-(1-7)] with dermorphin caused a great increase in opiate potency of the analog. [Dermorphin (1-7)]-beta c-EP was 120 times more potent than beta c-EP in the guinea pig ileum assay, 49 times more potent in the mouse vas deferens assay; and only 4 times more potent in the rat vas deferens assay. Replacement of NH2-terminal 1-13 segment of human beta-EP [beta h-EP-(1-13)] with dynorphin-A-(1-13) caused an increase in opiate potency in both the guinea pig ileum and rabbit vas deferens assays, a complete loss of potency in the rat vas deferens assay, and no change in the mouse vas deferens assay. In comparison with dynorphin-A-(1-13), the hybrid peptide was less potent in the guinea pig ileum assay as well as in mouse and rabbit vas deferens assay. It is suggested that beta c-EP-(8-31) facilitates the dermorphin moiety to act on opiate mu and delta receptors but not on the epsilon receptor, while beta h-(14-31) reduces the action of dynorphin on mu, delta and kappa receptors.

Amino Acid Sequence↗