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K W Li

Publications and source records attributed to K W Li.

At least 55 records · Page 3Linked to original sources
PubMed1995

Identification of a novel amide peptide, GLTPNMNSLFF-NH2, involved in the control of vas deferens motility in lymnaea stagnalis.

Peptide messengers are crucial for the execution of male copulatory processes in the snail Lymnaea stagnalis. In this study, we report the purification of a novel peptide from the penis complex of L. stagnalis. This peptide enhances the contraction frequency and contraction amplitude of the vas deferens. Amino acid sequencing and mass spectrometry indicate that the primary structure is GLTPNMNSLFF-NH2. We propose that this amide peptide modulates the transfer rate of semen in the vas deferens.

Amino Acid Sequence↗
PubMed1995

Co-localized neuropeptides conopressin and ALA-PRO-GLY-TRP-NH2 have antagonistic effects on the vas deferens of Lymnaea.

We examined functional aspects of co-localization of neuropeptides involved in the regulation of male copulation behaviour in the simultaneous hermaphrodite snail Lymnaea stagnalis. The copulation behaviour is controlled by several types of peptidergic neurons that include a cluster of neurons in the anterior lobe of the right cerebral ganglion. All anterior lobe neurons express the gene encoding Ala-Pro-Gly-Trp-NH2 (APGWamide), and a subset of neurons also express the vasopressin-related conopressin gene. Immunocytochemical and peptide chemical experiments show that both APGWamide and conopressin are transported to the penis complex and the vas deferens via the penis nerve. Co-localization of the two peptides was also observed in some, but not all, axon bundles that run along the vas deferens. APGWamide and conopressin were structurally identified from the penis complex with vas deferens. Conopressin excites the vas deferens in vitro, whereas APGWamide inhibits the excitatory effects of conopressin, both in a dose-dependent fashion. We propose that the antagonistic effects of these peptides on the vas deferens underlie its peristalsis. Thus, these peptides play an important role in the control of ejaculation of semen during copulation.

Amino Acid Sequence↗
PubMed1994

Direct peptide profiling by mass spectrometry of single identified neurons reveals complex neuropeptide-processing pattern.

A novel strategy combining peptide fingerprinting of single neurons by matrix-assisted laser desorption ionization mass spectrometry, molecular cloning, peptide chemistry, and electrospray ionization mass spectrometry was used to study the intricate processing pattern of a preprohormone expressed in identified neurons, the neuroendocrine light yellow cells (LYCs) of the gastropod mollusc, Lymnaea stagnalis. The cDNA encoding the precursor, named prepro-LYCP (LYCPs, light yellow cell peptides), predicts a straightforward processing into three peptides, LYCP I, II, and III, at conventional dibasic processing sites flanking the peptide domains on the precursor. However, matrix-assisted laser desorption ionization mass spectrometry of single LYCs revealed trimmed variant peptides derived from LYCP I and II. The variants were much more abundant than the intact peptides, indicating that LYCP I and II serve as intermediates in a peptide-processing sequence. Using the molecular masses of the peptides as markers to guide their isolation by well established purification methods, the structural identities of the peptides could be confirmed by amino acid sequencing. Furthermore, matrix-assisted laser desorption ionization mass spectrometry could detect colocalization of a novel peptide with the LYCPs.

Amino Acid Sequence↗
PubMed1994

Structural identification, neuronal synthesis, and role in male copulation of myomodulin-A of Lymnaea: a study involving direct peptide profiling of nervous tissue by mass spectrometry.

We used a strategy combining immunodetection, peptide chemistry, and a novel method, direct peptide fingerprinting of neurons and small pieces of nerve by using matrix-assisted laser desorption ionization mass spectrometry, to structurally identify and localize the neuropeptide myomodulin-A in the mollusc, Lymnaea stagnalis. Lymnaea myomodulin appeared to be identical to Aplysia myomodulin-A and is produced by many central neurons, including neurons located in the ventral lobe of the right cerebral ganglion that innervate the penis complex via the penis nerve. Myomodulin-A could also be characterized from the penis complex, and physiological concentrations of the peptide enhanced the relaxation rate of electrically induced contractions of the penis retractor muscle in vitro in a dose-dependent fashion.

Animals↗
PubMed1994

Processing, axonal transport and cardioregulatory functions of peptides derived from two related prohormones generated by alternative splicing of a single gene in identified neurons VD1 and RPD2 of Lymnaea.

The VD1/RPD2 mRNA precursor in identified neurons VD1 and RPD2 of the freshwater snail Lymnaea stagnalis is alternatively spliced to yield two related variants encoding two distinct yet related preprohormones, named the VD1/RPD2-A and -B preprohormones. Here, we report the isolation and structural characterization of alpha 1, alpha 2 and beta peptides from dissected neurons VD1 and RPD2. The alpha 1 and alpha 2 peptides are derived from VD1/RPD2-A and B prohormones, respectively, whereas beta peptide is identical for both prohormones. In addition, we report the isolation and structural characterization of the alpha 2 peptide from the heart, demonstrating that the mature peptides are transported and released in the heart. The pharmacological actions of synthetic alpha 1 and alpha 2 peptides on isolated auricle preparations of the Lymnaea heart were examined. The two alpha peptides have similar excitatory effects on beat rate and beat amplitude, while their potencies differed considerably, indicating that alternative splicing results in structurally and functionally overlapping, through non-identical, sets of peptides.

Amino Acid Sequence↗
PubMed1994

Neuropeptide expression and processing as revealed by direct matrix-assisted laser desorption ionization mass spectrometry of single neurons.

Neuropeptides were directly detected in single identified neurons and the neurohemal area of peptidergic (neuroendocrine) systems in the Lymnaea brain by using matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). The samples were placed in matrix solution and ruptured to allow mixing of cell contents with the matrix solution. After formation of matrix crystals, the analytes were analyzed by MALDI-MS. It was surprising that clean mass spectra were produced, displaying extreme sensitivity of detection. In one of the neuroendocrine systems studied, we could demonstrate for the first time, by comparing the peptide patterns of soma and of neurohemal axon terminals, that processing of the complex prohormone expressed in this system occurs entirely in the soma. In the other system studied, novel peptides could be detected in addition to peptides previously identified by conventional molecular biological and peptide chemical methods. Thus, complex peptide processing and expression patterns could be predicted that were not detected in earlier studies using conventional methods. As the first MALDI-MS study of direct peptide fingerprinting in the single neuron, these experiments demonstrate that MALDI-MS forms a new and valuable approach to the study of the synthesis and expression of bioactive peptides, with potential application to single-cell studies in vertebrates, including humans.

Animals↗
PubMed1994

Structural analysis of underivatized and derivatized aminophospholipids and phosphatidic acid by positive ion liquid secondary ion and collisionally induced dissociation tandem mass spectrometry.

This paper describes an approach for structurally analyzing aminophospholipids, including glycerophosphatidylethanolamine, glycerophosphatidylserine and their lyso analogues, and glycerophosphatidic acid by positive-ion liquid secondary-ion and four-sector tandem mass spectrometry. Polar head groups and the composition and position of the fatty acid chains in aminophospholipid species as well as diacyl and monoacyl phosphatidylethanolamine and their plasmalogen species can be characterized by collisionally induced dissociation of [M+H]+ ions of underivatized molecular species. Tandem mass spectrometry of [M+Na]+ and [M + 2Na-H]+ ions of 9-fluorenylmethyloxycarbonyl-derivatized glycerophosphatidylethanolamine and glycerophosphatidylserine induces fragmentation processes, but product ions are generally less structurally informative. Collisionally induced dissociation of [M + 2Na-H]+ ions of the molecular species of glycerophosphatidic acid not only yields information on the composition and position of fatty acid chains, but also allows the double bond location to be identified. This common positive-ion method is particularly effective in characterizing the molecular species of natural ethanolamine glycerophospholipids because of the advantage of a low detection limit. This approach also represents a useful alternative for the analysis of the molecular species from natural aminophospholipids by tandem mass spectrometry.

Amines↗
PubMed1994

Processing and targeting of a molluscan egg-laying peptide prohormone as revealed by mass spectrometric peptide fingerprinting and peptide sequencing.

The neuroendocrine cerebral caudodorsal cells of Lymnaea stagnalis initiate and coordinate ovulation and egg mass production and associated behaviors through the release of a complex set of peptides that are derived from the caudodorsal cell hormone-I (CDCH-I) precursor. We have previously characterized the CDCH-I peptide. In the present study, we isolated and amino acid sequenced by conventional peptide chemistry five additional peptides, epsilon-peptide, calfluxin, alpha-caudodorsal cell peptide, delta-peptide, and carboxyl-terminally located peptide, from the cerebral commissure, the neurohemal area of the caudodorsal cells. Fingerprinting by matrix-assisted laser desorption mass spectrometry of peptides in the commissure demonstrated the presence of all sequenced peptides and, in addition, could identify two other peptides derived from pro-CDCH-1, the beta 1- and beta 3-peptides. These findings together with previous immunocytochemical studies enabled us to define cleavage sites and major processing events of pro-CDCH-1. Pro-CDCH-1 is initially cleaved in the Golgi apparatus into carboxyl- and amino-terminal parts, each of which is sorted into distinct vesicle classes that traffic to different intracellular sites. As a result, in the commissure, peptides derived from the carboxyl-terminal part, including CDCH-1, are present at a many-fold higher concentration than those derived from the amino-terminal part.

Amino Acid Sequence↗
PubMed1994

Genomic organization of the FMRFamide gene in Lymnaea: multiple exons encoding novel neuropeptides.

Based on the sequencing of genomic and cDNA clones, we were able to determine that the FMRFamide gene consists of five exons covering at least 20 kb and predict the presence of further novel peptides. The exons are alternatively spliced: exon I (hydrophobic leader sequence) to exon II (tetrapeptides) and exon I to exons III (heptapeptides), IV, and V. A cDNA clone encoding the heptapeptides is described and has also been shown to encode further novel peptides SKPYMRFamide, HDYMRFamide, and SSFPRYamide. Analysis of the right internal parietal nerve using mass spectrometry showed that the novel peptide SKPYMRFamide was cleaved from the precursor. This peptide excites neurons, suggesting a physiological function in the CNS.

Amino Acid Sequence↗
PubMed1993

Purification and characterization of a novel tetradecapeptide that modulates oesophagus motility in Lymnaea stagnalis.

The isolated oesophagus of Lymnaea stagnalis was used as a bioassay preparation to identify peptides that modulate the activity of the oesophagus. Several modulatory messengers were detected from the oesophageal extract. One of these, having an excitatory action, was further purified from both the oesophagus and the central nervous system through high performance gel permeation chromatography followed by serial reversed phase high performance liquid chromatography. The purified peptide was chemically characterized through amino acid sequencing and mass spectrometry as GFRANSASRVAHGY-NH2.

Amino Acid Sequence↗
PubMed1993

Amino acid sequence of the small cadmium-binding protein (MP II) from Nereis diversicolor (annelida, polychaeta). Evidence for a myohemerythrin structure.

The primary sequence of the low-molecular-mass cadmium-binding protein metalloprotein II of Nereis diversicolor (Hediste diversicolor, recent denomination) has been determined. This protein is composed of 119 amino acids and has 80.8% identity with the N. diversicolor myohemerythrin [Takagi, T. & Cox, J. A. (1991) FEBS Lett. 285, 25-27]. The fact that iron, which normally binds to myohemerythrin, is not found to be associated with the cadmium-binding protein metalloprotein II in cadmium-exposed animals could be the result of the complete abolition of the iron-binding capacity of the protein due to the binding of cadmium.

Amino Acid Sequence↗
PubMed1993

Processing of the FMRFamide precursor protein in the snail Lymnaea stagnalis: characterization and neuronal localization of a novel peptide, 'SEEPLY'.

In the pulmonate snail Lymnaea stagnalis, FMRFamide-like neuropeptides are encoded by a multi-exon genomic locus which is subject to regulation at the level of mRNA splicing. We aim to understand the post-translational processing of one resulting protein precursor encoding the tetrapeptide FMRFamide and a number of other putative peptides, and determine the distribution of the final peptide products in the central nervous system (CNS) and periphery of Lymnaea. We focused on two previously unknown peptide sequences predicted by molecular cloning to be encoded in the tetrapeptide protein precursor consecutively, separated by the tetrabasic cleavage site RKRR. Here we report the isolation and structural characterization of a novel non-FMRFamide-like peptide, the 22 amino acid peptide SEQPDVDDYLRDVVLQSEEPLY. The novel peptide is colocalized with FMRFamide in the CNS in a number of identified neuronal systems and their peripheral motor targets, as determined by in situ hybridization and immunocytochemistry. Its detection in heart excitatory motoneurons and in nerve fibres of the heart indicated that the novel peptide may play a role, together with FMRFamide, in heart regulation in the snail. The second predicted peptide, STEAGGQSEEMTHRTA (16 amino acids), was at very low abundance in the CNS and was only occasionally detected. Our current findings, suggestive of a distinct pattern of post-translational processing, allowed the reassessment of a previously proposed hypothesis that the two equivalent sequences in the Aplysia FMRFamide gene constitute a molluscan homologue of vertebrate corticotrophin releasing factor-like peptides.

Amino Acid Sequence↗
PubMed1993

Idiopathic calcification of the basal ganglia.

Idiopathic calcification of the basal ganglia is a rare disorder characterized by neuropsychiatric abnormalities, a movement disturbance of parkinsonian and/or choreoathetoid type and dense calcification of the basal ganglia. We report a 60 year old patient diagnosed as having delusional disorder and tardive dyskinesia who was subsequently found to be suffering from idiopathic calcification of the basal ganglia.

Basal Ganglia↗
PubMed1993

Postoperative pulmonary tissue changes following autogenous lung and artificial lung bypass. Comparative studies.

With regard to pulmonary tissue damage secondary to extracorporeal circulation, most studies focused on the pulmonary histological changes after artificial lung bypass, but none compared the pre- and post-operative pulmonary changes of artificial lung with those of autogenous lung. We compared the changes in autogenous lung oxygenation and artificial lung oxygenation during cardiopulmonary bypass (CPB). The results showed that autogenous lung oxygenation is less injurious to the lung than bubble oxygenation and is more physiological. We think that during open heart surgery with autogenous lung bypass, the lung is functioning physiologically and supplied with normal amount of oxygen, while using bubble oxygenation for CPB, the native lung, the only nonperfused organ, is insulted by pulmonary ischemia, which is one of the most important reasons for pulmonary injury after CPB.

Animals↗
PubMed1992

Two gonadotropin-releasing hormones from African catfish (Clarias gariepinus).

Two forms of gonadotropin-releasing hormone (GnRH) have been purified from brain extracts of the African catfish, Clarias gariepinus, using reverse-phase high performance liquid chromatography (HPLC) and radioimmunoassay (RIA). The amino acid sequences of both forms of African catfish GnRH were determined using Edman degradation after digestion with pyroglutamyl aminopeptidase. In addition, both GnRHs were studied by mass spectrometry. The primary structure of African catfish GnRH I is identical to Thai catfish GnRH I, pGlu-His-Trp-Ser-His-Gly-Leu-Asn-Pro-Gly-NH2, and the primary structure of African catfish GnRH II is identical to the widely distributed and highly conserved chicken GnRH II, pGlu-His-Trp-Ser-His-Gly-Trp-Tyr-Pro-Gly-NH2.

Amino Acid Sequence↗
PubMed1992

The Light Green Cells of Lymnaea: a neuroendocrine model system for stimulus-induced expression of multiple peptide genes in a single cell type.

We review recent experiments showing that the cerebral neuroendocrine Light Green Cells (LGCs) of the freshwater snail, Lymnaea stagnalis, express a family of distinct though related molluscan insulin-related peptide (MIP) genes. The LGCs are involved in the regulation of a wide range of interrelated life processes associated with growth, (energy) metabolism and reproduction. We consider the mechanism of generation of diversity among MIPs, and present evidence that conditions with distinct effects on growth, metabolism and reproduction also can induce distinct patterns of expression of the MIP and schistosomin genes. The stimulus-dependent expression of multiple neuropeptide genes enormously increases the adaptive potential of a peptidergic neuron. We suggest that this contributes significantly to the information-handling capacity of the brain.

Amino Acid Sequence↗
PubMed1992

Isolation and chemical characterization of a novel insulin-related neuropeptide from the freshwater snail, Lymnaea stagnalis.

A novel molluscan insulin-related peptide (MIP) III, has been isolated from alcohol extracts of the neurohaemal area of the cerebral neuroendocrine light-green neurones of Lymnaea stagnalis. MIP III was purified by sequential high-performance gel-permeation chromatography followed by reverse-phase HPLC. MIP III is a heterodimer connected by disulphide bonds. Edman degradation analysis and subsequent alignment with the A and B chains of the previously identified MIP I and II showed that the 24-amino-acid peptide with the sequence pQSRPSIVC(E)CCFNQCTVQ(E)LLAYC represents the MIP III A chain, and the 37-amino-acid peptide sequence TTQHTCSILSRPHPRGLCGSTLANMVQWLCSTYTTSS the B chain. The overall amino acid sequence of MIP III shows about 50% similarity with those of MIP I and II, and only 20-40% similarity with other peptides of the insulin superfamily. Important structural features, e.g. disulphide bridges and the hydrophobic core, are conserved in MIP III.

Amino Acid Sequence↗