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Biomedical subjects

C C Chang

Publications and source records attributed to C C Chang.

At least 19 recordsLinked to original sources
PubMed1995

Regulation and immunolocalization of acyl-coenzyme A: cholesterol acyltransferase in mammalian cells as studied with specific antibodies.

Acyl-coenzyme A:cholesterol acyltransferase (ACAT) catalyzes the formation of intracellular cholesterol esters in various tissues. We recently reported the cloning and expression of human macrophage ACAT cDNA. In the current study, we report the production of specific polyclonal antibodies against ACAT by immunizing rabbits with the recombinant fusion protein composed of glutathione S-transferase and the first 131 amino acids of ACAT protein. Immunoblot analysis showed that the antibodies cross-reacted with a 50-kDa protein band from a variety of human cell lines. These antibodies immunodepleted more than 90% of detergent-solubilized ACAT activities from six different human cell types, demonstrating that the 50-kDa protein is the major ACAT catalytic component in these cells. In multiple human tissues examined, the antibodies recognized protein bands with various molecular weights. These antibodies also cross-reacted with the ACAT protein in Chinese hamster ovary cells. Immunoblot analysis showed that the ACAT protein contents in human fibroblast cells, HepG2 cells, or Chinese hamster ovary cells were not affected by sterol in the medium, demonstrating that the main mechanism for sterol-dependent regulation of ACAT activity in these cells is not change in ACAT protein content. As revealed by indirect immunofluorescent microscopy, the ACAT protein in tissue culture cells was located in the endoplasmic reticulum. This finding, along with earlier studies, suggests that cholesterol concentration in the endoplasmic reticulum may be the major determinant for regulating ACAT activity in the intact cells.

Animals↗
PubMed1995

cDNA sequence analysis and expression of alpha-bungarotoxin from Taiwan banded krait (Bungarus multicinctus).

The cDNA encoding alpha-bungarotoxin (alpha-BuTx) was constructed from the cellular RNA isolated from the venom glands of Bungarus multicintus by RT-PCR. The primers deduced from the N-, C-terminal amino acid residues and the conserved middle segment of the sequence, i.e., R25KMWC29 in alpha-BuTx could specifically amplify cDNAs encoding amino acid residues from 1 to 29 and 25 to 74 of alpha-BuTx. The complete cDNA was obtained by linking sufficient amounts of the two cDNA fragments by PCR. The amino acid sequence translated from the alpha-BuTx cDNA sequence was identical to that of native toxin. The PCR product was then cloned into pGEX vector and transformed in DH5 E. coli strain. The fused protein expressed in host strain exhibited the same antigenicity as native alpha-BuTx. This communication is the first report for the cloning and expression of a long alpha-neurotoxin from the venom glands.

Amino Acid Sequence↗
PubMed1995

Tissue-specific expression and cholesterol regulation of acylcoenzyme A:cholesterol acyltransferase (ACAT) in mice. Molecular cloning of mouse ACAT cDNA, chromosomal localization, and regulation of ACAT in vivo and in vitro.

Acyl-coenzyme A:cholesterol acyltransferase (ACAT) catalyzes the esterification of cholesterol with long chain fatty acids and is believed to play an important part in the development of atherosclerotic lesions. To facilitate the study of ACAT's role in this process, we have used the human ACAT K1 clone previously described (Chang, C. C. Y., Huh, H. Y., Cadigan, K. M. and Chang, T. Y. (1993) J. Biol. Chem. 268, 20747-20755) to isolate mouse ACAT cDNA from a liver cDNA library. The 3.7-kilobase cDNA clone isolated contains a 1620-base pair open reading frame which encodes a protein of 540 amino acids. The predicted mouse ACAT protein is 87% identical to the protein product of human ACAT K1 and shares many of the same secondary structural features, including two transmembrane domains, a leucine heptad motif consistent with dimer or multimer formation, and five regions homologous to the "signature sequences" found in other enzymes that catalyze acyl adenylation followed by acyl thioester formation and acyl transfer. Using the cDNA as a hybridization probe, we mapped the gene encoding mouse ACAT to chromosome 1 in a region syntenic to human chromosome 1 where the ACAT gene is located. Northern blot analysis and RNase protection assays of mouse tissues revealed that ACAT mRNA is expressed most highly in the adrenal gland, ovary, and preputial gland and is least abundant in skeletal muscle, adipose tissue, heart, and brain. To study the dietary regulation of ACAT mRNA expression in mouse tissues, we fed C57BL/6J mice a high-fat, high-cholesterol (HF/HC) atherogenic diet for 3 weeks and measured ACAT mRNA levels in various tissues by RNase protection. The HF/HC diet had little effect on ACAT mRNA levels in the small intestine, aorta, adrenal, or peritoneal macrophages, whereas hepatic ACAT mRNA levels were doubled in mice fed the atherogenic diet. ACAT activity in liver microsomes was similarly increased in cholesterol-fed mice, suggesting that mouse ACAT is regulated at least in part at the level of mRNA abundance. Additionally, a significant positive correlation was observed between ACAT activity and microsomal free cholesterol levels in chow- and cholesterol-fed mice, supporting the concept of cholesterol availability as a regulator of ACAT. To further investigate the regulation of ACAT activity under controlled conditions, ACAT-deficient Chinese hamster ovary cells were stably transfected with the mouse ACAT cDNA clone driven by a cytomegalovirus promoter. Two transfected Chinese hamster ovary cell lines that expressed the mouse ACAT transgene regained the ability to esterify cholesterol.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1995

A novel receptor involved in T-cell activation.

Optimal T-cell activation and T-cell expansion require triggering by T-cell antigen receptors and co-stimulatory signals provided by accessory cells. A major co-stimulatory pathway involves crosslinking the CD28 molecule on T cells by its ligands CD80 or CD86 expressed on antigen-presenting cells. But recent studies on CD28-deficient mice have indicated that CD28 is not required for all T-cell responses and that additional T-cell co-stimulatory pathways exist. Here we describe a novel glycoprotein, of relative molecular mass 70,000 (M(r) 70K), designated SLAM, that belongs to the immunoglobulin gene superfamily, which is involved in T-cell stimulation. SLAM is constitutively expressed on peripheral-blood CD45ROhigh memory T cells, T-cell clones, immature thymocytes, and a proportion of B cells, and is rapidly induced on naive T cells after activation. Engagement of SLAM enhances antigen-specific proliferation and cytokine production by T cells carrying the CD4 antigen (CD4+). Particularly, the production of interferon-gamma (IFN-gamma) is strongly upregulated, even in T helper type 2 (Th2) CD4+ T-cell clones, whereas no induction of interleukin (IL)-4 or IL-5 production was observed in Th1 clones. In addition, the engagement of SLAM induces directly the proliferation of CD4+ T-cell clones and preactivated T cells, in the absence of any other stimuli, and without CD28 involvement. Thus SLAM is a novel receptor on T cells that, when engaged, potentiates T-cell expansion in a CD28-independent manner and induces a Th0/Th1 cytokine production profile.

Amino Acid Sequence↗
PubMed1995

Developmental changes in the expression of gamma-aminobutyric acidA/benzodiazepine receptor subunit mRNAs in the murine inferior olivary complex.

The pharmacological and physiological properties of ligand-gated ion channels are dependent on their subunit composition; spontaneously occurring changes in subunit composition during neuronal development may result in dramatic functional differences between embryonic and adult forms of the receptor complex. In the present study, in situ hybridization with antisense cRNA probes was used to examine the subunit composition of the gamma-aminobutyric acidA/benzodiazepine (GABAA/BZ) receptor in the developing inferior olivary complex. This receptor is thought to be a pentameric chloride channel comprised of selected alpha, beta, gamma, delta, and rho subunits, the majority of which have several isoforms: alpha 1-6, beta 1-4, gamma 1-4, and rho 1,2. Among the 13 subunit variants present in the mammalian central nervous system, alpha 2-5, beta 3, and gamma 1,2 mRNAs are expressed at significant levels in the inferior olivary complex. Two clearly different temporal patterns of GABAA/BZ receptor subunit mRNA expression were observed: The expression of alpha 3, alpha 5, beta 3, and gamma 2 mRNAs was at a peak during embryonic and early postnatal development followed by rapid down-regulation thereafter. Conversely, alpha 2, alpha 4, and gamma 1 mRNA expression was very low or absent during early development, and a pronounced increase was observed at the end of postnatal week 1. These studies suggest that there are developmental changes in the subunit composition of the GABAA/BZ receptor in inferior olivary neurons. These changes in subunit expression, which occur during a period of major alterations in afferent and efferent synaptic connections, may subserve a change in the role of GABA from its function as a neurotrophic factor to that of an inhibitory neurotransmitter.

Animals↗
PubMed1995

Physiological and regenerative acetylcholine release from motor nerve: differential inhibitions by vesamicol and omega-agatoxin IVA.

Stimulation of mammalian motor neurons can elicit Ca(2+)-dependent regenerative release of acetylcholine and prolonged endplate depolarization when the enzymatic degradation of the neurotransmitter is inhibited. Unlike physiological phasic release of acetylcholine, the regenerative release is sensitive to L-type Ca2+ channel blockers. We studied the effects of vesamicol (an inhibitor of active transport of acetylcholine into synaptic vesicles) and omega-agatoxin IVA (a blocker of the motor nerve P-type Ca2+ channel) on these two types of acetylcholine release to compare the vesicle pools and Ca2+ channels responsible for the release. When coupled with repetitive stimulations, vesamicol decreased mean amplitude of miniature endplate potentials, resulting in a skewed distribution to lower amplitude, reduced quantal content of endplate potentials and decreased immediate available pool of acetylcholine. omega-Agatoxin IVA had no effect on miniature endplate potential but inhibited quantal content of endplate potential. The mean inhibitory concentration was around 5-10 nM. Vesamicol and omega-agatoxin IVA decreased the probability of triggering regenerative release. However, the magnitude and duration of regenerative release, once triggered, were not depressed by either agent. It appears that the majority of Ca2+ necessary for regenerative release is translocated via omega-agatoxin IVA-insensitive Ca2+ channels, which can be activated by prolonged depolarization of nerve terminals induced by accumulated acetylcholine. The results suggest that different Ca2+ channels are activated in the regenerative (L-type) and phasic (P-type) acetylcholine release, which utilize different pools of synaptic vesicles.

Acetylcholine↗
PubMed1995

The structural loop II of cobrotoxin is the main binding region for nAChR and epitope in the region is conformation-dependent.

Modification of positively charged residues, Lys and Arg, in cobrotoxin revealed that Lys-27, Lys-47, Arg-28, Arg-30, Arg-33, and Arg-36 of cobrotoxin were essential for the lethality and binding activity to nicotinic acetylcholine receptor (nAChR). The antigenicity of cobrotoxin was drastically diminished when Lys-47, Arg-28, Arg-30, Arg-33, and Arg-36 were modified, while that of the Lys-27-modified derivative was not significantly changed. The CD spectra of cobrotoxin displayed similar patterns after modification of Lys-27, Lys-47, and Arg-28. These findings suggest that Lys-27, Lys-47, and Arg-28 residues may be related to the direct binding to nAChR, and that there is no involvement of Lys-27 in the antigenic determinants of cobrotoxin. Extending the modification to Arg-30, Arg-33, and Arg-36 caused progressive conformational changes of cobrotoxin and resulted in decreased binding activity to antibody and nAChR. This indicates that Arg-30, Arg-33, and Arg-36 may be of structural importance for maintaining the active conformation of cobrotoxin. These results, together with the facts that Tyr-25, Tyr-35, and Trp-29 of cobrotoxin are important in nAChR binding activity, but Trp-29 and Tyr-35 residues are not essential for the antigenicity, suggest that the structural loop II of cobrotoxin is the main binding region for nAChR and the epitope in that region is conformation-dependent.

Amino Acid Sequence↗
PubMed1995

Analysis of a conformation-independent epitope and a conformational epitope in a protein: a study on cobrotoxin from Taiwan cobra venom.

The antibodies against cobrotoxin were separated into two antibody preparations by successive affinity chromatographies on reduced and S-carboxymethylated (RCM)-cobrotoxin-Sepharose, and cobrotoxin-Sepharose columns. The antibodies (abbreviated as Abcf-i) that bound with the RCM-cobrotoxin-Sepharose were verified to specifically recognize the continuous epitopes of cobrotoxin, which were insensitive to conformational changes. Whilst the antibodies (abbreviated as Abcf-d) that did not bind with the RCM-cobrotoxin-Sepharose column recognized the conformational epitopes in cobrotoxin. The two antibody preparations were employed to screen the antigenic peptides derived from the proteolytic hydrolysate of cobrotoxin and RCM-cobrotoxin. Five antigenic peptides (AP-4, AP-5, AP-10, AP-11, and AP-12) were obtained from the acid protease A-digested hydrolysate of cobrotoxin, and two antigenic peptides (V8-2 and V8-4) were found in the hydrolysate of RCM-cobrotoxin after hydrolysis with Saccharomyces aureus V8 protease. The segments at positions 1-21 and 22-38 encompassed the peptide fractions, AP-4, AP-5, V8-2, and V8-4, that reacted with Abcf-i, indicating that the two segments bore the continuous epitopes of cobrotoxin. Alternatively, AP-10, AP-11, and AP-12 reacted with both Abcf-i and Abcf-d. The structures of the three peptides had a common segment at positions 43-62, suggesting that this region comprised the conformation-independent epitopes as well as conformational epitopes in cobrotoxin. These results reflected that the conformation-independent and conformational epitopes in a protein can be separately identified.

Amino Acid Sequence↗
PubMed1995

Determination of soluble nAChR-binding activity of alpha-neurotoxins by an innovative precipitation with DEAE-Sephacel.

A simple and convenient method for determining the binding activity of alpha-neurotoxins toward soluble nicotinic acetylcholine receptor (snAChR) by precipitation with DEAE-Sephacel was established. The determination was carried out by incubation of 125I-neurotoxin with snAChR, followed by precipitation with DEAE-Sephacel. The DEAE-Sephacel particles bind negatively charged snAChR with high affinity and simultaneously precipitate the 125I-neurotoxin bound to the receptors. After centrifugation, the free 125I-neurotoxin in the supernatant was counted, and the amounts of neurotoxins bound to snAChR could be determined. Two alpha-neurotoxins, cobrotoxin and alpha-bungarotoxin, were employed to verify the feasibility of this determination. The different binding characteristics of cobrotoxin and alpha-bungarotoxin to snAChR could be distinguished. This method required only small quantities of DEAE-Sephacel (7 mg), snAChR (0.54 micrograms), and 125I-neurotoxin (90 fmol) for each reaction, and minimized the handling of isotopic materials as compared with the conventional methods. This method is reliable, reproducible, and superior to current methods for the determination of the snAChR-binding activity for alpha-neurotoxins.

Binding, Competitive↗
PubMed1995

Vasodilator agents modulate rat glomerular mesangial cell growth and collagen synthesis.

Mesangial cell activation and extracellular matrix accumulation are hallmarks of many forms of glomerulonephropathy. We investigated the effect of several agents possessing vasodilating activities on rat mesangial cell growth and collagen synthesis. Using cell counting and a modified MTT assay, it was shown that dipyridamole, pentoxifylline, dicentrine, prazosin and doxazosin all caused a dose-dependent inhibition of serum-stimulated rat mesangial cell proliferation. Platelet-derived growth factor-induced cell proliferation was also inhibited by doxazosin and pentoxifylline. Dipyridamole and pentoxifylline inhibited collagen synthesis in confluent mesangial cells while dicentrine and doxazosin did not. The procollagen alpha 1 (I) mRNA expression was also decreased by dipyridamole and pentoxifylline. These results suggested that, in addition to dipyridamole, pentoxifylline and alpha 1-adrenoceptor blockers may have a potential to delay the progression of chronic glomerulopathy associated with mesangial proliferation.

Analysis of Variance↗
PubMed1995

Evidence showing a different repertoire of antibodies against unfolded cobrotoxin in anticobrotoxin and anti-reduced and S-carboxymethylated cobrotoxin antisera.

Antibodies which specifically reacted with reduced and S-carboxy-methylated(RCM) cobrotoxin were purified from anticobrotoxin antisera and anti-RCM-cobrotoxin antisera, respectively. Results using a competitive immunoassay revealed that the antibodies from anti-RCM-cobrotoxin antisera had a greater affinity for RCM-cobrotoxin than for cobrotoxin. Whereas the reactivity of the antibodies from the anticobrotoxin antisera toward cobrotoxin and RCM-cobrotoxin had a reversed order of binding. In contrast to observations made with S. aureus V8 protease-digest hydrolysates, the antigenic structures of RCM-cobrotoxin recognized by antibodies from anti-RCM-cobrotoxin antisera were notably affected following hydrolysis of RCM-cobrotoxin with chymotrypsin. Moreover, the chymotryptic hydrolysates showed a comparable reactivity as RCM-cobrotoxin toward the antibodies purified from anticobrotoxin antibodies, but a decrease in antigenicity with the V8 protease hydrolysates. These results reveal that the repertoire of antibodies against the unfolded cobrotoxin are not the same in anticobrotoxin and anti-RCM-cobrotoxin antisera. Moreover, it suggests that the repertoire of antibodies from the different sources against the same antigen can be differentiated by measurement of their reactivities with the proteolytic hydrolysates of the antigen.

Animals↗
PubMed1995

Power spectral analysis of systemic arterial pressure signals during open heart surgery.

BACKGROUND: The close relationship between the balance of sympathetic and parasympathetic tones and the results of power spectral analysis (PSA) of systemic arterial pressure (SAP) has been suggested recently. The purpose of this study was to further describe the changes of balance between the two autonomic nervous components during open heart surgery for coronary arterial disease (CAD) or valvular heart disease (VHD) with the PSA technique of SAP. METHODS: By relaying the SAP signals to a personal computer utilizing a power spectral analysis algorithm, radial arterial pressure signals of 27 patients (11 with CAD for coronary artrial bypass graft, as CAD groups; 16 with VHD, as VHD group, including 8 for aortic valve replacement and 8 for mitral valve replacement) during open heart surgery were monitored in a continuous, on-line and real-time manner. On-line power spectral analysis was performed according to the five different stages of the operation. RESULTS: Power density tended to increase in the high frequency (HF) band during the ventilator-supported stages of the open heart procedures, i.e., the pre-cardiopulmonary bypass (CPB) and the off-CPB periods. The power density in other frequency bands during the whole course of general anesthesia otherwise decreased significantly, with the lowest values occurring during CPB. The power density in very low frequency (VLF) band was much higher than in other bands during CPB, and became the major component of total power density in this period. CONCLUSIONS: The balance between sympathetic and parasympathetic components of autonomic nervous system changes rapidly during open heart surgery and needs careful monitoring. By utilizing PSA of SAP, mathematical error might be an obstacle of using LF:HF ratio as an index of autonomic balance during the CPB period when the HF density approaches zero.

Aged↗
PubMed1994

High-density multiplex detection of nucleic acid sequences: oligonucleotide ligation assay and sequence-coded separation.

We describe a non-isotopic, semi-automated method for large-scale multiplex analysis of nucleic acid sequences, using the cystic fibrosis transmembrane regulator (CFTR) gene as an example. Products of a multiplex oligonucleotide ligation assay (OLA) are resolved electrophoretically from one another and from unligated probes under denaturing conditions with fluorescence detection. One ligation probe for each OLA target carries a fluorescent tag, while the other probe carries an oligomeric non-nucleotide mobility modifier. Each OLA product has a unique electrophoretic mobility determined by the ligated oligonucleotides and the mobility-modifier oligomer arbitrarily assigned (coded) to its target. The mobility range for practical mobility modifiers is much wider than the accessible range from unmodified ligated oligonucleotides of practical length. Each mobility modifier is built from phosphoramidite monomers in a stepwise manner on its associated oligonucleotide using an automated synthesizer. The resulting mobility modifiers lower the probe-target duplex Tm by less than 3 degrees C and retard probe-target annealing by less than 50%, with negligible effect on OLA yield and specificity. This method is especially useful for allelic discrimination in highly polymorphic genes such as CFTR.

Autoanalysis↗
PubMed1994

A nested case-control study on association between hepatitis C virus antibodies and primary liver cancer in a cohort of 9,775 men in Taiwan.

Most studies on the association between antibodies against hepatitis C virus (anti-HCV) and primary liver cancer (PLC) were limited to case-series, or cross-sectional case-control studies leaving a controversy on causal temporality. A nested case-control study on 38 newly-developed PLC patients and 152 matched controls selected from a cohort of 9,775 men in Taiwan recruited from September, 1984, to February, 1986, was carried out to examine the relation between HCV infection and PLC. Case-control pairs were matched on age (+/- 1 year), residence, and the date at recruitment. Serum samples collected from study subjects at the initial recruitment were examined for anti-HCV by enzyme immunoassay and hepatitis B surface antigen (HBsAg) by reverse passive hemagglutination assay combined with radioimmunoassay. History of cigarette smoking, alcohol consumption, vegetable consumption, vegetarian habit, and chronic liver diseases were also obtained through standardized interviews according to a structured questionnaire at the recruitment. After adjusting for HBsAg status and other risk factors, the anti-HCV was significantly associated with the development of PLC showing a multivariate-adjusted relative risk of 88.24. The results suggest that HCV infection may play an important role in the etiology of human PLC in Taiwan.

Adult↗
PubMed1994

BCL-6 and the molecular pathogenesis of B-cell lymphoma.

The results presented identify the first genetic lesion associated with DLCL, the most clinically relevant form of NHL. Although no proof yet exists of a role for these lesions in DLCL pathogenesis, the feature of the BCL-6 gene product, its specific pattern of expression in B cells, and the clustering of lesions disrupting its regulatory domain strongly suggest that deregulation of BCL-6 expression may contribute to DLCL development. A more precise definition of the role of BCL-6 in normal and neoplastic B-cell development is the goal of ongoing study of transgenic mice engineered either to express BCL-6 under heterologous promoters or lacking BCL-6 function due to targeted deletions. In addition to contributing to the understanding of DLCL pathogenesis, the identification of BCL-6 lesions may have relevant clinical implications. DLCL represent a heterogeneous group of neoplasms which are treated homogeneously despite the fact that only 50% of patients experience long-term disease-free survival (Schneider et al. 1990). The fact that BCL-6 rearrangements identify biologically and clinically distinct subsets of DLCL suggests that these lesions may be useful as markers in selection of differential therapeutic strategies based on different risk groups. Furthermore, the BCL-6 rearrangements can be used to identify and monitor the malignant clone with sensitive PCR-based techniques. Since clinical remission has been observed in a significant fraction of DLCL cases, these markers may serve as critical tools for sensitive monitoring of minimal residual disease and early diagnosis of relapse (Gribben et al. 1993).

Animals↗
PubMed1993

Brequinar sodium, mycophenolic acid, and cyclosporin A inhibit different stages of IL-4- or IL-13-induced human IgG4 and IgE production in vitro.

We investigated the effect of cyclosporin A (CsA), mycophenolic acid (MPA), and brequinar sodium (BQ) on human IgG4 and IgE synthesis induced by IL-4 or IL-13. BQ inhibited IL-4 and IL-13-induced IgG4 and IgE synthesis in cultures of peripheral blood mononuclear cells (PBMC) or highly purified B cells costimulated by anti-CD40 mAbs in a dose-dependent fashion. CsA and MPA had either suppressive or enhancing effects depending on the concentrations tested. Interestingly, BQ inhibited IgG4 and IgE synthesis at concentrations of 10(-6)-10(-8) M, which did not affect T or B cell proliferation, indicating that the inhibitory effects of BQ on Ig production were not directly related to inhibition of T or B cell proliferation. In contrast, the inhibitory effects of MPA on Ig production were directly associated with inhibitory effects on T and B cell proliferation. CsA blocked T and B cell proliferation at the same concentration (10(-7) M) which enhanced IgG4 and IgE synthesis, indicating that reduction in T or B cell proliferation correlated with enhanced IgE production. CsA also inhibited CD40 ligand expression and IL-2, IL-4, IL-5, IFN-gamma, and GM-CSF production by activated CD4+ T cell clones, whereas MPA and BQ were ineffective, indicating that these compounds do not inhibit early events in T cell activation. Collectively, our data indicate that BQ, MPA, and CsA block different stages of the IgG4 and IgE production process. In addition, we observed that CsA and MPA, in contrast to BQ, at lower concentrations can also have potentiating effects on the production of these Ig isotypes.

Antigens, CD↗
PubMed1992

Chemical modification of arginine residues in alpha-bungarotoxin.

The reaction of alpha-bungarotoxin (alpha-BuTX) with 1,2-cyclohexanedione resulted in the modification of only Arg-72 but arginine at position 36 or 72, as well as both were modified by reaction of the toxin with p-hydroxyphenylglyoxal. No derivative modified at Arg-25 was obtained, indicating that this residue may be located in the interior region of alpha-BuTX molecule. Monoderivative at Arg-72 showed about 50% of the lethal toxicity and binding activity of alpha-BuTX to nicotinic acetylcholine receptor (AChR), while the activity was decreased to one-third when the invariant Arg-36 was modified, indicating that the latter residue is more closely related to the interaction of the toxin with AChR. Approx. 13% of the residual activity was observed when both arginine residues at 36 and 72 were modified. The antigenicity of alpha-BuTX was still retained essentially intact after Arg-36 or -72 was modified, whereas it decreased to 50% when both these arginine residues were modified. The present study indicates that Arg-36 and -72 in alpha-BuTX may be involved in the multipoint contact between the toxin and AChR, but neither is absolutely essential for the binding.

Amino Acids↗