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H M Li

Publications and source records attributed to H M Li.

At least 37 records · Page 2Linked to original sources
PubMed1996

Crystal structure of an acidic neurotoxin from scorpion Buthus martensii Karsch at 1.85 A resolution.

The crystal structure of an acidic scorpion neurotoxin, BmK M8, purified from Chinese scorpion Buthus martensii Karsch (BmK), has been determined by the molecular replacement method. It is the first structure of an acidic alpha-scorpion neurotoxin reported so far. The crystals adopt a symmetry of space group P2(1) and contain one molecule per asymmetric unit. The structure has been refined to an R factor of 18.1% using reflection data in the range of 8 to 1.85 A resolution, with standard deviations from ideal geometry of 0.017 A and 2.43 degrees for bond length and angle, respectively. The 12 residues at the C terminus with unknown sequence were determined by crystallographic refinement. The refined model shows that the structural core, consisting of a motif beta alpha beta beta, is similar to that of toxin II from Androctonus australis Hector (AaH II) or Variant 3 from Centruroides sculpturatus Ewing (CsE V3). The three conformationally variable loops protruding from this structural core are different from that of AaH II, and especially from that of CsE V3. Compared with the most potent and basic alpha-toxin AaH II, the BmK M8 is a relatively inactive toxin (1100 times less active than AaH II) with an unusually low isoelectric point (pI 5.3). Sequence alignment of the two toxins shows a difference of 26 residues (40.6%). Among them four basic or neutral residues in AaH II, namely Val10, Lys28, Val55 and Gly59, are changed to acidic glutamate in BmK M8. The residues Glu10, Glu28 and Glu55 of BmK M8 are located on a surface (Face B), opposite the "conserved hydrophobic surface" (Face A). The latter is a functionally important area proposed by Fontecilla-Camps et al. Our observations suggest that in addition to Face A, Face B may also be involved in the biological activity of scorpion toxins. The structure of BmK M8 shows an evident conformational change of the alpha-amino group at the N terminus and a deorganization of Arg2 caused by the mutation D53A. These structural changes may also be responsible for the weak toxicity of BmK M8. In association with the information from chemical modifications, a multisite binding mode for toxin-receptor interaction and three "toxic regions" in relevance to the binding process, including Face A, Face B and Site C, are proposed. Face A, mainly consisting of Tyr5, 35, 47, the alpha-amino group, Arg2 and Asp3, may be more essential for the binding. Face B, mainly comprising conserved residues Tyr14, 21, Lys28 and Val55, may contribute to the high efficacy of the binding process and substitutions by acidic residues in this area could strongly weaken the toxic activity. Site C, formed by Lys58 and Arg62 at the C terminus and Arg41 and Tyr42 from loop 38-44, may be involved in binding site specificity.

Amino Acid Sequence↗
PubMed1996

Protein targeting and integration signal for the chloroplastic outer envelope membrane.

Most proteins in chloroplasts are encoded by the nuclear genome and synthesized in the cytosol. With the exception of most quter envelope membrane proteins, nuclear-encoded chloroplastic proteins are synthesized with N-terminal extensions that contain the chloroplast targeting information of these proteins. Most outer membrane proteins, however, are synthesized without extensions in the cytosol. Therefore, it is not clear where the chloroplastic outer membrane targeting information resides within these polypeptides. We have analyzed a chloroplastic outer membrane protein, OEP14 (outer envelope membrane protein of 14 kD, previously named OM14), and localized its outer membrane targeting and integration signal to the first 30 amino acids of the protein. This signal consists of a positively charged N-terminal portion followed by a hydrophobic core, bearing resemblance to the signal peptides of proteins targeted to the endoplasmic reticulum. However, a chimeric protein containing this signal fused to a passenger protein did not integrate into the endoplasmic reticulum membrane. Furthermore, membrane topology analysis indicated that the signal inserts into the chloroplastic outer membrane in an orientation opposite to that predicted by the "positive inside" rule.

Amino Acid Sequence↗
PubMed1996

Flow-induced detachment of adherent platelets from fibrinogen-coated surface.

A study of the shear forces under which adherent platelets of different morphologies can be detached from surfaces was carried out using a newly designed tapered flow chamber, which covered the entire shear range of physiological circulation. Platelets that naturally settled on a fibrinogen-coated surface were exposed to shear flow and were subsequently processed for scanning electron microscopic observation. We found that 1) the density of platelets remaining after flow exposure decreased with local shear stress, 2) adherent platelets of different morphologies withstood different levels of shear stress: most round cells and 40% of the cells that had a few short pseudopods were detached at < 10 dyn/cm2, whereas most spread cells could withstand 50 dyn/cm2, 3) pulsatile flow was more effective in removing adherent platelets than equivalent steady flow, 4) cytochalasin D and colchicine retarded platelet shape change and made them more easily detached by shear forces, and 5) metabolic energy-depleted platelets spread readily and formed shear-resistant clumps. Our observations indicated that adherent platelets of different morphologies on a fibrinogen-coated surface could withstand different levels of flow shear stress.

Blood Circulation↗
PubMed1995

Signal-transduction pathways controlling light-regulated development in Arabidopsis.

All metazoan cells are able to make decisions about cell division or cellular differentiation based, in part, on environmental cues. Accordingly, cells express receptor systems that allow them to detect the presence of hormones, growth factors and other signals that manipulate the regulatory processes of the cell. In plants, an unusual signal-light-is required for the induction and regulation of many developmental processes. Past physiological and molecular studies have revealed the variety and complexity of plant responses to light but until recently very little was known about the mechanisms of those responses. Two major breakthroughs have allowed the identification of some photoreceptor signalling intermediates: the identification of photoreceptor and signal transduction mutants in Arabidopsis, and the development of single-cell microinjection assays in which outcomes of photoreceptor signalling can be visualized. Here, we review recent genetic advances which support the notion that light responses are not simply endpoints of linear signal transduction pathways, but are the result of the integration of a variety of input signals through a complex network of interacting signalling components.

Arabidopsis↗
PubMed1994

Formalin induced FOS-like immunoreactive neurons in the trigeminal spinal caudal subnucleus project to contralateral parabrachial nucleus in the rat.

By combining the retrograde-labeling method of injecting Fluoro-Gold (FG) into the parabrachial nucleus (PB) and the immunocytochemical staining of the FOS-like immunoreactive neurons (FLNs) in the trigeminal spinal caudal subnucleus (TSCS) induced by s.c. formalin injection into the perioral region in the rat, it was demonstrated that there are FLNs, FG-labeled neurons and neurons containing both FOS-like immunoreactivity and FG fluorescence in the TSCS. The three kinds of labeled neurons are distributed mainly in laminae I, II and V of the TSCS and there are also some neurons containing both FOS-like immunoreactivity and FG distributed in the adjacent ventrolateral reticular formation. The retrograde-labeling of FG- and double-labeled neurons showed contralateral predominance. In addition, we found that there are retrogradely labeled neurons in bilateral nuclei of the solitary tract with a contralateral predominance. The results suggest that FOS-like immunoreactivity might serve as an indicator for the nociceptive response after formalin injection into the trigeminal region and that the PB might be an important relay station for the further processing of the nociceptive information relayed from the trigeminal afferents. As the PB is known as a relay structure for visceral sensory pathway, it is proposed that there might be viscero-somatic convergence in this nucleus.

Animals↗
PubMed1994

Arabidopsis mutants define downstream branches in the phototransduction pathway.

Light regulates the development of Arabidopsis seedlings in a variety of ways, including inhibition of hypocotyl growth and promotion of leaf development, chloroplast differentiation, and light-responsive gene expression. Mutations that uncouple most or all of these responses from light control have been described, for example, det1, det2, and cop1. To identify regulatory components that define downstream branches in the light-regulated signal transduction pathway, mutants specifically affected in only one light-regulated response were isolated. A screen was designed to isolate mutants that overexpressed the CAB (photosystem II type I chlorophyll a/b-binding proteins) genes in the dark, by use of transgenic line containing a T-DNA construct with two CAB3 promoter-reporter fusions. Eight mutants that showed aberrant expression of both CAB3 promoters were isolated and were designated doc mutants (for dark overepression of CAB). All of the mutants have normal etiolated morphology in the dark. Genetic and phenotypic analyses indicate that most of the mutations are recessive and define at least three loci (doc1, doc2, doc3). Unlike det1 and det2 mutants, which affect the expression of CAB and RBCS (the small subunit of RuBP carboxylase) to approximately the same extent, all three doc mutations are much more specific in derepressing the expression of CAB. The phenotypes of doc mutants suggest that morphological changes can be genetically separated from changes in CAB gene expression. Moreover, the regulation of CAB gene expression can be separated further from the regulation of RBCS gene expression. Epistasis studies suggest that DOC1 and DET3 act downstream from DET1 on two separate branches in the phototransduction pathway. In contrast, DOC2 appears to act on a distinct pathway from DET1. Mutations in doc1, doc2, or doc3 also impair plant growth under short-day conditions.

Arabidopsis↗
PubMed1994

Molecular cloning of a chloroplastic protein associated with both the envelope and thylakoid membranes.

Chloroplasts consist of six morphologically distinct compartments. Each compartment has a specific set of polypeptides that perform distinct biochemical functions. We report here the identification of a membrane-associated protein with a novel localization. This protein was synthesized as a 37 kDa precursor and was processed to a mature protein of 30 kDa after being imported into isolated pea chloroplasts. Fractionation of chloroplasts showed that the 30 kDa mature protein was associated with both of the envelope membranes as well as with thylakoid membranes. Immunocyto-chemical localization of the 30 kDa protein revealed that the protein occurred in clusters in the vicinity of both the envelope and the thylakoid. Possible functions of this 30 kDa protein, inferred from its novel localization pattern, are discussed.

Amino Acid Sequence↗
PubMed1994

[Neuropsychiatric disturbances and neuro-electrophysiological examination in patients with arsenic poisoning].

Neuropsychiatric disturbances caused by arsenic poisoning in 28 patients were reported. The incidence was 32.2% of all the arsenic poisoning patients. The patients were classified into three types according to their clinical symptoms. The neurologic type constituted 46.4%, mental type 21.4% and mixed type 32.1% of the patient respectively. Various abnormalities were found in EEG, EMG, EP and EKG examination. EMG and EP were still abnormal in most of the patients after treatment for three months. The results showed that arsenic poisoning may cause neuropsychiatric disturbances. The symptoms in these patients are serious and the recovery is quite slow.

Adolescent↗
PubMed1994

[Cataract extraction and intraocular lens implantation in patients with uveitis].

Thirty two patients with uveitis had a posterior chamber intraocular lens implantation after extracapsular cataract extraction. After a mean follow-up period of 5.4 months (range 3 to 29 months), the corrected visual acuity in 93.7% of the patients reached to 0.5 or better. Postoperative complications included posterior capsule opacification and iris posterior synechiae. The results suggest that careful selection of candidates for IOL implantation be crucial for successful cataract surgery in uveitis patients.

Adolescent↗
PubMed1994

[Congenital cataract extraction with primary posterior chamber IOL implantation].

Congenital cataracts in 41 eyes were extracted with primary implantation of posterior chamber IOLs, of which 35 were the one-piece PMMA variety. The patients were followed up 3-28 months, averaging 9.5 months. Corrected visual acuities of 1.0-1.5 were achieved in 29.3% and 0.5 or better in 85.4% of the eyes. Postoperative complications were few. Among 34 eyes of preoperative low vision, visual acuities of 0.3 or over were restored to 31 eyes (91.2%) immediately after the operation.

Adolescent↗
PubMed1994

[Intraocular lens implantation in Fuchs' syndrome].

10 patients of Fuchs' syndrome underwent ECCE with IOL implantation. The etiopathology of this disease, the prevention and management of intra- and post-operative complications were discussed. Three months after the operation, the rate of corneal endothelial cell loss was 23.96%. After an average follow-up of 20 months, 80% of the cases obtained corrected visual acuity of 0.5 and over, indicating that the procedure had been rewarding and safe.

Adult↗
PubMed1993

Prenatal ontogeny of substance P-like immunoreactivity in the nucleus tractus solitarii and dorsal motor nucleus of the vagus nerve of the human fetus: an immunocytochemical study.

By using immunocytochemical method, the prenatal ontogeny of substance P-like immunoreactivity (SP-LI) was demonstrated in the dorsal motor nucleus of the vagus nerve (nX) and the nucleus tractus solitarii (nTS) of the human fetus at fetal age (menstruation age) of 11.5 weeks to 40 weeks. The time of initial appearance of SP-LI in the human brainstem nTS was between the fetal age 11.5 weeks and 16 weeks. At fetal age 16 weeks, the nTS showed moderate density of SP-LI fibers and terminals in subnucleus dorsalis of the nTS and nX. While the fetus grew, the density of SP-LI in the human fetus brainstem nTS and nX increased gradually from fetal age 16 weeks to 40 weeks. According to the Nissl staining, at fetal age 23 weeks, the nTS of human fetus can be subdivided into dorsal, medial, dorsolateral, ventrolateral, ventral and gelatinosus subnuclei. The cytoarchitectonic subdivisions of human fetus nTS is in good agreement with the results obtained by immunocytochemical staining. These findings indicated that substance P (SP) might play an important role in the development of human brainstem nX, nTS, their related cranial nerves, and in their functional establishment during the prenatal period.

Benzoxazines↗
PubMed1993

Regulation of gene expression by light.

The past year has seen significant advances in the biochemical, genetic and molecular dissection of the light signal transduction and developmental pathways that lead to photoregulated gene expression in higher plants. A major part of recent research has focused on the assignment of biological functions to the various photoreceptors, the genetic dissection of the photoreceptor action pathways, and the identification of the cis-acting sequences and trans-acting factors that regulate the downstream light-regulated genes.

Base Sequence↗
PubMed1992

Information for targeting to the chloroplastic inner envelope membrane is contained in the mature region of the maize Bt1-encoded protein.

Based on the protein sequence deduced from a cDNA clone, it has been proposed that the maize bt1 locus encodes an amyloplast membrane metabolite translocator protein (Sullivan, T. D., Strelow, L. I., Illingworth, C. A., Phillips, R. L., and Nelson, O. E., Jr. (1991) Plant Cell 3, 1337-1348). The present work provides further evidence for this hypothesis by showing that the gene product of Bt1 could be imported into chloroplasts in vitro and processed to lower molecular weight mature proteins. More importantly, the imported mature proteins were localized to the inner envelope membrane, where metabolite translocators are located in plastids. In addition, the location of information for targeting to the inner membrane was investigated by constructing and analyzing the import of chimeric precursor proteins. A chimeric protein with the transit peptide of the precursor to the small subunit of ribulose-1,5-bisphosphate carboxylase fused to the mature region of the Bt1-encoded protein was targeted to the inner envelope membrane of chloroplasts. Moreover, a chimeric protein with the transit peptide of the Bt1-encoded protein fused to the mature protein of the light-harvesting chlorophyll a/b binding protein was targeted to the thylakoid. These results indicate that the transit peptide of the Bt1-encoded protein functions primarily as a stromal targeting sequence. The information for targeting to the chloroplastic inner envelope membrane is contained in the mature region of the protein.

Base Sequence↗