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Y M Xiong

Publications and source records attributed to Y M Xiong.

9 recordsLinked to original sources
PubMed2001

Structure-function of the putative I-domain within the integrin beta 2 subunit.

The central region (residues 125-385) of the integrin beta(2) subunit is postulated to adopt an I-domain-like fold (the beta(2)I-domain) and to play a critical role in ligand binding and heterodimer formation. To understand structure-function relationships of this region of beta(2), a homolog-scanning mutagenesis approach, which entails substitution of nonconserved hydrophilic sequences within the beta(2)I-domain with their homologous counterparts of the beta(1)I-domain, has been deployed. This approach is based on the premise that beta(1) and beta(2) are highly homologous, yet recognize different ligands. Altogether, 16 segments were switched to cover the predicted outer surface of the beta(2)I-domain. When these mutant beta(2) subunits were transfected together with wild-type alpha(M) in human 293 cells, all 16 beta(2) mutants were expressed on the cell surface as heterodimers, suggesting that these 16 sequences within the beta(2)I-domain are not critically involved in heterodimer formation between the alpha(M) and beta(2) subunits. Using these mutant alpha(M)beta(2) receptors, we have mapped the epitopes of nine beta(2)I-domain specific mAbs, and found that they all recognized at least two noncontiguous segments within this domain. The requisite spatial proximity among these non-linear sequences to form the mAb epitopes supports a model of an I-domain-like fold for this region. In addition, none of the mutations that abolish the epitopes of the nine function-blocking mAbs, including segment Pro(192)-Glu(197), destroyed ligand binding of the alpha(M)beta(2) receptor, suggesting that these function-blocking mAbs inhibit alpha(M)beta(2) function allosterically. Given the recent reports implicating the segment equivalent to Pro(192)-Glu(197) in ligand binding by beta(3) integrins, these data suggest that ligand binding by the beta(2) integrins occurs via a different mechanism than beta(3). Finally, both the conformation of the beta(2)I-domain and C3bi binding activity of alpha(M)beta(2) were dependent on a high affinity Ca(2+) binding site (K(d) = 105 microm), which is most likely located within this region of beta(2).

Amino Acid Sequence↗
PubMed1999

A new antifungal peptide from the seeds of Phytolacca americana: characterization, amino acid sequence and cDNA cloning.

An antifungal peptide from seeds of Phytolacca americana, designated PAFP-s, has been isolated. The peptide is highly basic and consists of 38 residues with three disulfide bridges. Its molecular mass of 3929.0 was determined by mass spectrometry. The complete amino acid sequence was obtained from automated Edman degradation, and cDNA cloning was successfully performed by 3'-RACE. The deduced amino acid sequence of a partial cDNA corresponded to the amino acid sequence from chemical sequencing. PAFP-s exhibited a broad spectrum of antifungal activity, and its activities differed among various fungi. PAFP-s displayed no inhibitory activity towards Escherichia coli. PAFP-s shows significant sequence similarities and the same cysteine motif with Mj-AMPs, antimicrobial peptides from seeds of Mirabilis jalapa belonging to the knottin-type antimicrobial peptide.

Amino Acid Sequence↗
PubMed1999

Expression and purification of the BmK M1 neurotoxin from the scorpion Buthus martensii Karsch.

The gene encoding a neurotoxin (BmK M1) from the scorpion Buthus martensii Karsch was expressed in Saccharomyces cerevisiae at a high level with the alcohol dehydrogenase promoter. SDS-PAGE of the culture confirmed expression and showed secretion into medium from yeast. Recombinant BmK M1 was purified rapidly and efficiently by ion exchange and gel filtration chromatography to homogeneity, produced a single band on tricine-SDS-PAGE, and processed the homologous N-terminus. Amino acid analysis and N-terminal sequencing demonstrated that the recombinant toxin was processed correctly from the alpha-mating factor leader sequence and was chemically identical to the native form. The expressed recombinant BmK M1 was toxic for mice, which indicated that it was biologically active. Quantitative estimation showed that recombinant BmK M1 had an LD(50) similar to that of the native toxin.

Animals↗
PubMed1999

The cDNA sequence of an excitatory insect selective neurotoxin from the scorpion Buthus martensi Karsch.

The full-length cDNA of an excitatory insect selective neurotoxin was amplified from total cDNAs of venomous glands of the scorpion Buthus martensi Karsch (BmK) using the 3'RACE and 5'RACE (rapid amplification of cDNA ends, RACE) method and sequenced. The cDNA encoded a precursor of the insect toxin of 88 amino acid residues, including a signal peptide of 18 residues and a mature toxin of 70 residues. The cDNA deduced sequence of this toxin was homologous with the determined amino acid sequence of BmK IT1, an excitatory insect toxin purified from the scorpion venom, except for three different residues, two at the positions 24-25, and another in the COOH-terminus of the toxin. Among them the COO-terminal residue Gly in the cDNA deduced sequence was predominantly different from the conserved residue Asn found in other known scorpion excitatory insect toxins.

Amino Acid Sequence↗
PubMed1999

Molecular characterization of a new excitatory insect neurotoxin with an analgesic effect on mice from the scorpion Buthus martensi Karsch.

Besides the neurotoxins active on mammals, a new excitatory insect selective toxin with a mice analgesic activity was found and purified from the venom of the scorpion Buthus martensi Karsch (BmK) (Ji, Y.H., Mansuelle, P., Terakawa, S., Kopeyan, C., Yanaihara, N., Hsu, K., Rochat, H., 1996. Toxicon 34, 987; Luo, M.J., Xiong, Y.M., Wang, M., Wang, D.C., Chi, C.W., 1997. Toxicon 35, 723.). This peptide (designated as BmK IT-AP) is composed of 72 amino acid residues. Its primary structure was determined by automated Edman degradation of the N-terminal part of the reduced and S-carboxamidemethylated protein and its lysylendopeptidase degraded fragments. Based on the determined sequence, the gene specific primers were designed and synthesized for 3' and 5' RACE (rapid amplification of cDNA ends). Their partial cDNA fragments obtained by 3' and 5' RACEwere cloned and sequenced and the full length cDNA sequence of BmK IT-AP was then completed by overlapping their two partial cDNA sequences. It encodes a precursor of 90 amino acid residues: a signal peptide of 18 residues and a mature peptide of 72 residues which are consistent with the determined protein sequence of BmK IT-AP. The genomic DNA of the peptide was also amplified by PCR from the scorpion genomic DNA and sequenced, which is a first report on the genomic structure of a scorpion toxin specific for insects. Its sequence revealed an intron of 590 bp inserted in the end part of the signal peptide. The peptide caused a fast excitatory contraction paralysis on house fly larvae. Furthermore, the peptide also showed an obvious analgesic effect on mice, as assayed by using a twisting test model. This effect of BmK IT-AP well characterized at molecular level is first reported among the known scorpion insect neurotoxins.

Amino Acid Sequence↗
PubMed1997

Purification and sequence determination of a new neutral mammalian neurotoxin from the scorpion Buthus martensii Karsch.

A new neutral mammalian neurotoxin, designated BmK M4, with an isoelectric point (pI) of 7.6 and a relatively low toxicity (LD50 = 4.0 +/- 0.25 microgram/g mice, i.v.) was purified from the venom of scorpion Buthus martensii Karsch (BmK). The complete amino acid sequence of the toxin composed of 64 amino acid residues was determined by automated Edman degradation of the N-terminal part of the reduced and S-carboxamidomethylated protein (up to 30 amino acid residues) and its peptide fragments degraded by lysylendopeptidase or Staphylococcus aureus Va protease. The calculated mol. wt based on the amino acid composition was 7001. By comparison with the sequences of other basic BmK mammalian neurotoxins, it was concluded that the weaker toxicity and lower pI value of BmK M4 might be the result of mutations H10E, R18G and K28D. The sequence comparison of BmK M4 with an acidic toxin, BmK M8, showed that the weakest toxicity and acidic property of BmK M8 may be the consequence of mutations K8D, D53A, V55E and V59E. The substitution of 21 Gly in BmK M4 for Tyrin other BmK toxins may also be of importance. In their tertiary structures, these mutated charged residues are mainly distributed in the surface (face B) that is roughly opposite to the "conserved hydrophobic surface" (face A) proposed by Fontecilla-Camps et al. in 1982. Therefore the toxin-receptor interaction may take a multiposition mode.

Amino Acid Sequence↗
PubMed1997

The CDNA and genomic DNA sequences of a mammalian neurotoxin from the scorpion Buthus martensii Karsch.

The cDNA library of venomous glands of the scorpion Buthus martensii Karsch (BmK) was constructed. A cDNA encoding a mammalian neurotoxin corresponding to the known alpha-type toxin, BmK M1, was amplified by polymerase chain reaction (PCR) and cloned, and its full-length sequence was determined. The open reading frame encoded the precursor of BmK M1 with 84 amino acid residues, including a signal peptide of 19 residues, a mature toxin of 64 residues and an additional C-terminal residue Arg which might be cleaved off by proteinase postprocessing immediately after protein synthesis. Based on the determined cDNA sequence and using the total DNA of the scorpion as a template, the gene of BmK M1 was also amplified by PCR and sequenced. The genomic DNA sequence revealed an intron of 408 base pairs present within the signal peptide region. Both the intron and exon of BmK M1 share about 75% similarity with those of AaH I' another alpha-type mammalian neurotoxin in the scorpion Androctonus australis Hector.

Amino Acid Sequence↗
PubMed1991

Modulation of response to adenosine in vascular smooth muscle cells cultured in defined medium.

Cultured pig aortic smooth muscle cells maintain a viable, quiescent state in a chemically defined medium that contains 10(-6) M insulin, 5 micrograms/ml transferrin, and 0.2 mM ascorbate. DNA synthesis and DNA content were determined by measuring tritiated thymidine incorporation and DNA-binding to the fluorescent probe 4',6-diamidino-2-phenylindole, respectively. The majority of the population of cells in defined medium cultures were diploid. Tritiated thymidine uptake in cells in defined medium was one-tenth that observed in cells in fetal bovine serum-containing medium. The study of cellular cyclic AMP level in response to extracellular adenosine stimulation in dividing cells and quiescent cells showed that cells in defined medium had a lower extent of response to adenosine compared to cells cultured in serum-containing medium. Both the cell growth index and the response to adenosine of cells cultured in defined medium were reversible after replacing the medium with 10% fetal bovine serum-containing medium, which suggests that the cells in defined medium were healthy and were capable of modulating cellular metabolism depending on culture conditions.

Adenosine↗