Muscarinic Toxin-Like Proteins from Taiwan Banded Krait (Bungarus multicinctus) Venom: Purification, Characterization and Gene Organization

2002 ◽  
Vol 383 (9) ◽  
pp. 1397-1406 ◽  
Author(s):  
C. Chung ◽  
B.-N. Wu ◽  
C.-C. Yang ◽  
L.-S. Chang
Keyword(s):  
Amino Acid ◽  
Gene Organization ◽  
Receptor Subtype ◽  
Amino Acid Residues ◽  
Cd Spectra ◽  
Dna Encoding ◽  
Novel Proteins ◽  
Acrylamide Quenching ◽  
Venom Proteins ◽  
Quinuclidinyl Benzilate

Abstract Two novel proteins, BM8 and BM14, were isolated from Bungarus multicinctus (Taiwan banded krait) venom using the combination of chromatography on a SPSephadex C-25 column and a reversephase HPLC column. Both proteins contained 82 amino acid residues including 10 cysteine residues, but there were two amino acid substitutions at positions 37 and 38 (Glu37Ala38 in BM8; Lys37Lys38 in BM14). CD spectra and acrylamide quenching studies revealed that the gross conformation of BM8 and BM14 differed. In contrast to BM8, BM14 inhibited the binding of [3H]quinuclidinyl benzilate to the M2 muscarinic acetylcholine (mAchR) receptor subtype. Trinitrophenylation of Lys residues abolished the mAchRbinding activity of BM14, indicating that the Lys substitutions at positions 37 and 38 played a crucial role in the activity of BM14. The genomic DNA encoding the precursor of BM14 was amplified by PCR. The gene shared virtually identical structural organization with αneurotoxin and cardiotoxin genes. The intron sequences of these genes shared a sequence identity up to 84%, but the proteincoding regions were highly variable. These results suggest that BM8, BM14, neurotoxins and cardiotoxins may have originated from a common ancestor, and the evolution of snake venom proteins shows a tendency to diversify their functions.

Synthesis and conformation of sequential basic polypeptides with branched and linear side chains

1985 ◽  
Vol 50 (12) ◽  
pp. 2925-2936 ◽  
Author(s):  
Štěpánka Štokrová ◽  
Jan Pospíšek ◽  
Jaroslav Šponar ◽  
Karel Bláha
Keyword(s):  
Amino Acid ◽  
Salt Concentration ◽  
Salt Solution ◽  
Theoretical Work ◽  
Side Chain ◽  
Amino Acid Residues ◽  
Cd Spectra ◽  
Low Salt ◽  
Conformational Freedom ◽  
Basic Polypeptides

Polypeptides (Lys-X-Ala)n and (Lys-X-Gly)n in which X represents residues of isoleucine and norleucine, respectively, and polypeptide (Tle-Lys-Ala)n, were synthesized via polymerization of 1-hydroxysuccinimidyl esters of the appropriate tripeptides to complete previously studied series. Circular dichroism (CD) spectra of the respective polymers were measured as a function of pH and salt concentration of the medium. The results were correlated with those obtained previously with the same series containing different amino acid residues at the X-position. The helix forming ability of the polypeptides (Lys-X-Ala)n with linear X side chain was found to be independent of the length. In the series (Lys-X-Gly)n the unordered conformation was the most probable one except (Lys-Ile-Gly)n. This polymer assumed the β conformation even in low salt solution at neutral pH. An agreement with some theoretical work concerned with the restriction of conformational freedom of amino acid residue branching at Cβ atom with our experimental results is evident.

scholarly journals Molecular cloning, gene organization and expression of the human UDP-GalNAc:Neu5Acalpha2-3Galbeta-R beta1,4-N-acetylgalactosaminyltransferase responsible for the biosynthesis of the blood group Sda/Cad antigen: evidence for an unusual extended cytoplasmic domain

2003 ◽  
Vol 373 (2) ◽  
pp. 369-379 ◽  
Author(s):  
Maria-Dolores MONTIEL ◽  
Marie-Ange KRZEWINSKI-RECCHI ◽  
Philippe DELANNOY ◽  
Anne HARDUIN-LEPERS
Keyword(s):  
Amino Acid ◽  
Catalytic Domain ◽  
Human Gene ◽  
Short Form ◽  
Cytoplasmic Tail ◽  
Gene Organization ◽  
Sialic Acid Residue ◽  
Chromosome 17 ◽  
Soluble Form ◽  
Amino Acid Residues

The human Sda antigen is formed through the addition of an N-acetylgalactosamine residue via a β1,4-linkage to a sub-terminal galactose residue substituted with an α2,3-linked sialic acid residue. We have taken advantage of the previously cloned mouse cDNA sequence of the UDP-GalNAc:Neu5Acα2-3Galβ-R β1,4-N-acetylgalactosaminyltransferase (Sda β1,4GalNAc transferase) to screen the human EST and genomic databases and to identify the corresponding human gene. The sequence spans over 35 kb of genomic DNA on chromosome 17 and comprises at least 12 exons. As judged by reverse transcription PCR, the human gene is expressed widely since it is detected in various amounts in almost all cell types studied. Northern blot analysis indicated that five Sda β1,4GalNAc transferase transcripts of 8.8, 6.1, 4.7, 3.8 and 1.65 kb were highly expressed in colon and to a lesser extent in kidney, stomach, ileum and rectum. The complete coding nucleotide sequence was amplified from Caco-2 cells. Interestingly, the alternative use of two first exons, named E1S and E1L, leads to the production of two transcripts. These nucleotide sequences give rise potentially to two proteins of 506 and 566 amino acid residues, identical in their sequence with the exception of their cytoplasmic tail. The short form is highly similar (74% identity) to the mouse enzyme whereas the long form shows an unusual long cytoplasmic tail of 66 amino acid residues that is as yet not described for any other mammalian glycosyltransferase. Upon transient transfection in Cos-7 cells of the common catalytic domain, a soluble form of the protein was obtained, which catalysed the transfer of GalNAc residues to α2,3-sialylated acceptor substrates, to form the GalNAcβ1-4[Neu5Acα2-3]Galβ1-R trisaccharide common to both Sda and Cad antigens.

Melanin Concentrating Hormone Analogs: Contraction of the Cyclic Structure. III. CD Spectroscopic Study

1992 ◽  
Vol 57 (3) ◽  
pp. 614-620
Author(s):  
Ivo Frič ◽  
Michal Lebl ◽  
Victor J. Hruby
Keyword(s):  
Amino Acid ◽  
Spectroscopic Study ◽  
Spectral Properties ◽  
Membered Ring ◽  
Helical Conformation ◽  
Amino Acid Residues ◽  
Cyclic Structure ◽  
Cd Spectra ◽  
Melanin Concentrating Hormone ◽  
Hormone Analogs

A comparison of the CD spectra of MCH analogs differing in length but containing a heterodetic ring of the same size (compounds I, IV and VII or III, VI and IX) reveals that a conformational change occurs upon elongating the peptide chain from thirteen to seventeen amino-acid residues. The 5-17 fragments appear to prefer a β-turn conformation, whereas the 1-17 full-sequence peptides prefer α-helical conformation. Peptides containing seventeen-membered ring exhibit greater conformational adaptability (the incorporation of their cyclic moiety into an ordered conformation being easier) than those containing a twenty-six-membered ring. Spectral properties of the twenty-three-membered heterodetic ring in peptides II and V indicate that they do not possess highly ordered conformation.

scholarly journals Gene Organization, Expression, and Localization of Ribotoxin-Like Protein Ageritin in Fruiting Body and Mycelium of Agrocybe aegerita

2020 ◽  
Vol 21 (19) ◽  
pp. 7158
Author(s):  
Ilaria Baglivo ◽  
Sara Ragucci ◽  
Paolo D’Incecco ◽  
Nicola Landi ◽  
Rosita Russo ◽  
...  
Keyword(s):  
Amino Acid ◽  
Fruiting Body ◽  
Laser Scanning ◽  
Edible Mushroom ◽  
Gene Organization ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Amino Acid Residues ◽  
Agrocybe Aegerita ◽  
Fungal Genes

The edible mushroom Agrocybe aegerita produces a ribotoxin-like protein known as Ageritin. In this work, the gene encoding Ageritin was characterized by sequence analysis. It contains several typical features of fungal genes such as three short introns (60, 55 and 69 bp) located at the 5′ region of the coding sequence and typical splice junctions. This sequence codes for a precursor of 156 amino acids (~17-kDa) containing an additional N-terminal peptide of 21 amino acid residues, absent in the purified toxin (135 amino acid residues; ~15-kDa). The presence of 17-kDa and 15-kDa forms was investigated by Western blot in specific parts of fruiting body and in mycelia of A. aegerita. Data show that the 15-kDa Ageritin is the only form retrieved in the fruiting body and the principal form in mycelium. The immunolocalization by confocal laser scanning microscopy and transmission electron microscopy proves that Ageritin has vacuolar localization in hyphae. Coupling these data with a bioinformatics approach, we suggest that the N-terminal peptide of Ageritin (not found in the purified toxin) is a new signal peptide in fungi involved in intracellular routing from endoplasmic reticulum to vacuole, necessary for self-defense of A. aegerita ribosomes from Ageritin toxicity.

Conformation of branched polypeptides based on poly(L-lysine): The effect of terminal amino acids in the branches

1985 ◽  
Vol 50 (1) ◽  
pp. 228-244 ◽  
Author(s):  
Hana Votavová ◽  
Ferenc Hudecz ◽  
Judit Kajtár ◽  
Jaroslav Šponar ◽  
Karel Bláha ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Ionic Strength ◽  
Glutamic Acid ◽  
Amino Acid Residues ◽  
Ordered Structure ◽  
Cd Spectra ◽  
Helix Formation ◽  
Terminal Amino ◽  
Α Helix

CD Spectra of branched polypeptides based on poly(L-lysine) and containing three DL-alanine residues and one to three other L- or D-amino acid residues in the branches were measured in water, water-methanol and water-trifluoroethanol mixtures. In aqueous solutions dependence of the CD spectra on pH and ionic strength was studied. The effect of branch elongation was followed mainly with compounds containing glutamic acid. One terminal D-amino acid residue and also an extension by two L- or D-amino acid residues does not hinder the α-helix formation in the backbone but affects the conditions of its formation. In polypeptides with three L- or D-amino acids additional α-helical segments in the branches are assumed to be formed. For branches with L-amino acids the CD curves express additively the contributions of both helical components, in the case of D-amino acids the increasing population of the ordered structure in branches is manifested by compensation of dichroic contribution of the L-amino acid backbone leading even to enantiomorphous curves.

scholarly journals Biometric Estimation for Understanding the Nature of Vorticella Stalk Contraction-Extension Repeated Consequential Cyclic Processes

2021 ◽  
Vol 11 (4) ◽  
Author(s):  
Verma Dr. Amit Kumar
Keyword(s):  
Amino Acid ◽  
Data Bank ◽  
Diagnostic Tools ◽  
Amino Acid Residues ◽  
Novel Proteins ◽  
Different Types ◽  
Potential Applications ◽  
Cyclic Processes ◽  
Pharmaceutical Industries ◽  
Experimental Trials

Vorticella stalk is the storehouse of two types of novel proteins, known with the names of spasmins and batonnets. On the basis of nature of proteins and the arrangements of amino acid residues of these proteins, the repeated consequential cyclic processes of contraction dynamics worked by neutralizing the negatively charged amino acid residues as per the laws obeyed by Heber-Weiss, Nernst and Fenton reactions. H+-integrated physiological performances in combination with pCa (partial pressure of calcium ion concentrations) and DNFB (2,4 – dinitrofluorobenzene/Sanger’s reagent) concentration gradients at the range of 1mM to 5mM represented velocity inclination in acidic medium which were more actively pronounced if it was compared with alkaline medium where permeabilized stalk exaggerated potential biochemical-shift-perturbation if it was in respect of non-permeabilized live specimens in both artificial as well as in natural medium in different experimental trials under controlled electro-physiological instrumental setup conditions. On the basis of these experimental designs it was confirmed that spasmins and batonnets are two different types of novel proteins with multitudes of potential applications in the favour of biomedical engineering devices formulation, then their construction at the nano-scale where H+-integrated pCa dependent electrophysiological nature of recommended proteins were found more ROS (reactive oxygen species) resistant if there was the introduction of DNFB in fixed concentrations than in the acto-myosin as well as tubulin-dynein systems being exclusively controlled under post-translational biochemical reactions catalysed in the light of software based modern bioinformatics’ tools and techniques. In live as well as permeabilized specimens, different types of biochemical reaction kinetics of amino acid residues were performed at different rates among the sequentially determined spasmins and batonnets like novel proteins where molecular orientations and motive-force generation in measurable parameters per millisecond confirmed the electrophysiological significance of Vorticella stalks’ on the basis of colligative nature of novel proteins of saccular compartments of spasmoneme, the well explained active contractile organelle of the stalk in relation with other resembling proteins found in Protein Data Bank (PDB) as centrins, calmodulins and others significantly pronounce their life saving and medicinal properties. This present statement/study was aimed to know the biochemical behaviour of spasmins and batonnets like novel proteins in the light of electrochemical behavior of the Vorticella stalk under some selective chemical stress conditions, that’s why this research helped us to know the ROS resistance properties of novel protein polymers found in stalk. On the basis of which reference proteins as described in this paper can be used as a diagnostic tools in pharmaceutical industries in the favour of molecular medicines and drugs’ designing.

Conformation of branched polypeptides based on poly(L-lysine): Circular dichroism study

1980 ◽  
Vol 45 (3) ◽  
pp. 941-949 ◽  
Author(s):  
Hana Votavová ◽  
Ferenc Hudecz ◽  
Judit Kajtár ◽  
Mária Szekerke ◽  
Jaroslav Šponar ◽  
...  
Keyword(s):  
Circular Dichroism ◽  
Amino Acid ◽  
Main Chain ◽  
Side Chain ◽  
Amino Acid Residues ◽  
Cd Spectra ◽  
Circular Dichroïsm

Circular dichroism (CD) spectra of branched (multichain) polypeptides based on poly(L-lysine) with DL-alanine and other amino-acid residues in the side chain have been measured under various conditions of pH. The spectra were interpreted using spectra of poly(L-lysine) as standard. No straightforward correlation between polypeptide CD spectra and their structure could be detected. The contribution of the side chain CD to that of the main chain is additive in some cases, but in other cases qualitatively different CD spectra arise. The reason for this CD behavior is discussed.

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