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Biomedical subjects

H Pang

Publications and source records attributed to H Pang.

At least 91 records · Page 5Linked to original sources

The N terminus of human myelin basic protein consists of C2, C4, C6, and C8 alkyl carboxylic acids.

Peptide 1-21, generated by cyanogen bromide cleavage of each of two highly purified components of human myelin basic protein, components 1 and 8, gave a series of peaks in the fast atom bombardment mass spectra with m/z 2299, 2327, 2355, 2383, and 2411, indicating additions of 42, 70, 98, 126, and 154 atomic mass units respectively with m/z 2327 and 2355 as the dominant species. The pentafluorobenzyl esters prepared from an acid hydrolysate analyzed by negative ion chemical ionization gas chromatography mass spectrometry confirmed that C6, C8, and C10 fatty acids were present. These data demonstrated (i) that the N terminus of a myelin basic protein is not simply acetylated but contains C2, C4, C6, C8, and C10 fatty acids with C4 and C6 as the dominant species, (ii) the two components studied (C-1 and C-8) showed different relative amounts of C2 and C8 in particular, and (iii) human myelin basic protein is the first protein to be reported with a complex N terminus consisting of several alkyl carboxylic acid species.

Amino Acid Sequence↗

Binding of verocytotoxin 1 to its receptor is influenced by differences in receptor fatty acid content.

Globotriaosylceramide [(Gal alpha 1-4Gal beta 1-4Glc-ceramide (Gb3)] was separated from human kidney, and the fatty acid composition was determined. Semisynthetic Gb3 molecular species of corresponding fatty acid chain length were prepared and compared for verotoxin (VT) binding affinity by TLC overlay, and a quantitative binding assay was performed in the presence of auxiliary lipids. Our results indicate that, within the natural range, fatty acid chain length has little effect on verotoxin binding but that Gb3 molecular species containing different fatty acids can interact to provide a higher affinity toxin receptor than any of the individual component receptor species. Receptor function as assayed by TLC overlay was not always found to correlate with binding in a lipid environment. Short-chain fatty acid Gb3 molecular species could not function as VT receptors under these conditions. Evidence is presented to suggest that fatty acid chain length can have a stereoselective effect on carbohydrate conformation.

Adult↗

Genetic polymorphism of human factor H (HF, beta 1H globulin) in Chinese Han population in northeast China.

The distribution of human factor H of serum phenotypes were studied using ultrathin polyacrylamide gel isoelectric focusing (PAGIEF) and subsequent immunoblotting techniques in 203 Chinese of Han population in Liaoning Province of northeast China. The gene frequencies of HF*A and HF*B were 0.4828 and 0.5172, respectively. All the observed numbers of the phenotypes were in agreement with the expected numbers under the Hardy-Weinberg equilibrium. The gene frequencies among Chinese, Japanese, and Caucasian populations were compared.

China↗

Surface components of chylomicrons from rats fed glyceryl or alkyl esters of fatty acids: minor components.

The lipid class, fatty acid and molecular species composition of the minor polar surface components of rat lymph chylomicrons were determined during absorption of menhaden oil and corn oil or of the corresponding fatty acid ethyl esters. In addition to the previously reported minor polar lipids (sphingomyelin, phosphatidylserine, phosphatidylinositol, phosphatidic acid and lysophosphatidylcholine), we identified phosphatidylglycerol, dimethylphosphatidylethanolamine, ceramide and cholesteryl sulfate in the chylomicrons from both oil and ester feeding. The dietary fatty acids were found to be incorporated to a variable extent into the different phospholipid classes, the proportions of which remained the same during both types of feeding. No evidence was obtained for the presence of the minor glycerophospholipids characteristic of the lysosomal membranes (e.g., bis-phosphatidic, lysobisphosphatidic and semilysobis-phosphatidic acids), although special efforts were made to identify them. These results indicate that the chylomicrons arising from the monoacylglycerol and phosphatidic acid pathways of triacylglycerol biosynthesis become enveloped in closely similar monolayers of phospholipids. Hence, all triacylglycerols may be secreted from the villus cells via a common mechanism as suggested by the previously demonstrated convergence (at the 2-monoacylglycerol stage) of the monoacylglycerol and the phosphatidic acid pathways of mucosal triacylglycerol formation [Yang, Y.L., and Kuksis, A. (1991) J. Lipid Res. 32, 1173-1186].

Animals↗

Serological diversity and chemical structures of Campylobacter jejuni low-molecular-weight lipopolysaccharides.

Low-Mr lipopolysaccharides (LPS) of Campylobacter jejuni reference strains for serotypes O:1, O:4, O:23, and O:36 were examined through the liberation of core oligosaccharides by mild acid cleavage of the ketosidic linkage of 3-deoxy-D-manno-2-octulosonic acid residues to the lipid A moiety. The liberated oligosaccharides were examined for chemical structure by compositional analysis and methylated linkage analysis in conjunction with fast atom bombardment-mass spectrometry of permethylated oligosaccharide derivatives. The results showed (i) that the LPS contained short oligosaccharide chains of branched nonrepetitive structure, to many of which N-acetylneuraminic acid residues remained attached by 2----3 linkages to 4-linked D-galactose residues in the core structure; (ii) that serotypical differences, which are not readily defined through qualitatively similar compositions, are clearly reflected in variations in linkage types and sequences of sugar residues in the outer core attached to an inner region of invariable structure; but (iii) that the presence or absence of NeuAc residues does not appear to be a basis for serotypical differences. The results also showed that oligosaccharide chains from LPS of serotypes O:1 and O:4 are distinctly different and are distinct again from those of the cross-reacting serotypes O:23 and O:36, between whose core oligosaccharide chains no differences were found. It is concluded that the structurally variable low-Mr LPS from C. jejuni show greater similarities to the lipooligosaccharides from Neisseria spp. than to the highly conserved core regions of Salmonella species. Those strains (serotypes O:23 and O:36) which also furnish high-Mr LPS are unique among gram-negative bacteria in possessing both low-Mr molecules of the Neisseria lipooligosaccharide type and high-Mr LPS of the Salmonella smooth type.

Campylobacter jejuni↗

Genetic polymorphism of complement component C6, C7 and C8(1) in Chinese Han population in northeast China.

Distributions of complement phenotypes, C6, C7, and C8(1) were studied using thin agarose gel isoelectric focusing (AGIEF) or ultra-thin polyacrylamide gel isoelectric focusing (PAGIEF) and subsequent immunoblotting techniques in 203 Chinese Han population in Liaoning Province of northeast China. The gene frequencies were as follows: C6*A 0.4704, C6*B 0.5049, C6*B2 0.0148, C6*B3 0.0049, and C6*M 0.0049; C7*1 0.8251, C7*2 0.1108, C7*3 0.0320, and C7*4 0.0320; C8(1)*A 0.5567 and C8(1)B 0.4433, respectively. All the observed numbers of the phenotypes were in agreement with the expected numbers under the Hardy-Weinberg equilibrium. The gene frequencies among Chinese subpopulations and other various populations were compared.

Asian People↗

Mechanism and pathway of penicillopepsin-catalyzed transpeptidation and evidence for noncovalent trapping of amino acid and peptide intermediates.

Penicillopepsin acting on Nph-Ala2-amide (where Nph = p-nitrophenylalanyl) catalyzes a transpeptidation reaction which leads to the formation of Nph2-Ala2-amide, which arises from condensation of the substrate with enzyme-bound Nph, as the first product released from the enzyme. This is followed by a stage during which Nph3 and Ala2-amide are the major products. A small amount of Nph4 is also formed during this time. Nph and Nph2, formed during the reactions, are tightly, but probably not covalently, bound to the enzyme. They appear as free products only as a result of the cleavage of Nph3 and Nph4 and after most of the substrate Nph-Ala2-amide has been used up. They act as acceptors for the substrate and for Nph2-Ala2-amide. Nph3-Ala2-amide, formed by condensation of Nph-Ala2-amide or of Nph2-Ala2-amide with enzyme-bound Nph2 or Nph, respectively, is also released but is cleaved rapidly to give Nph3 and Ala2-amide. Incorporation of 18O from [18O]water into the carbonyl oxygens of the products is extensive and shows that release of the intermediates is slower than peptide bond cleavage and peptide bond formation. Hence the rate-limiting step in these reactions is product release. No 18O is incorporated into the initial substrate. We propose that Nph and Nph2 as intermediates are held in the active site by hydrogen bonds and by two strong electrostatic interactions.

Amino Acids↗

Diagnostic methods for the determination of iduronic acid in oligosaccharides.

A high-performance liquid chromatography (HPLC) method with pulsed amperometric detection (PAD) was used for the determination of the acid hydrolysis products of L-iduronic acid containing oligosaccharides isolated from biological sources. This HPLC-PAD method was compared with gas chromatographic (GLC) methods. Since acid hydrolysis of oligosaccharides can produce a number of products, several uronic acid derivatives were prepared by chemical synthesis. These well characterized standards in conjunction with mass spectrometry allowed for the identification of most of the products of methanolysis or hydrolysis of glycosamino-glycans, which included chondroitin sulfates A and B (dermatan sulfate), heparin, and hyaluronic acid. (4 M) HCl in methanol 100 degrees C for 24 h was found to be optimum for GLC and 1 M aqueous HCl for 4 h at 100 degrees C for HPLC-PAD. All of the monosaccharides, hexosamines, and uronic acids could be separately identified in a single chromatographic step using either technique. Good resolution, high sensitivity (low microgram samples) and rapid analysis makes these methods particularly useful for the determination of small amounts of glycosaminoglycans and other glycoconjugates found in samples isolated from biological sources. These two techniques are specifically designed to allow the qualitative determination of the carbohydrate content and composition of samples whose carbohydrate composition and content is completely unknown.

Chondroitin Sulfates↗

Synthesis and structural characterization of a cardioactive biotinylated digoxin analogue.

We have biotinylated the terminal glycose of digoxin by reaction of the periodate-oxidized steroid with biotin hydrazide. A biotinylated product (BD-1) was formed which retained significant digoxin receptor (Na+/K+ ATPase) binding activity. Sustained reaction resulted in a second biotinylated product (BD-2) which showed reduced receptor binding activity. The products were characterized by FAB mass spectroscopy and shown to be the mono- and di-biotinylated digoxin conjugates of the oxidized glycose moiety. These analogues may prove useful in determining the subcellular site of digoxin binding.

Biotin↗

Purification, characterization, and structural elucidation of the active moiety of the previously called "suppressor activating factor (SAF)".

Upon extensive purification of the serum-free supernatant produced by a mutant T cell line (6T-CEM), an immunosuppressive activity was found to reside in an oxidized product of spermine, spermine dialdehyde (SDA). The activity was purified to homogeneity from a serum-free supernatant by using gel filtration chromatography and reverse-phase C18 HPLC. Fast Atom Bombardment (FAB) mass spectral analysis revealed its MW to be 202 and Electron Impact (EI) analysis of the acetylated material identified the purified molecule to be spermine. In the presence of human or rodent plasma, spermine exhibited no immunosuppressive activity up to 2 mg/ml. However, when assayed in the presence of FCS, which contains polyamine oxidase (PAO), spermine is oxidized to its corresponding dialdehyde which is active at 0.1 microM/ml. We have previously described a high molecular weight suppressor activating factor (SAF) found in the serum-containing supernatant of the 6T-CEM cell line. Our preliminary biological data suggest that SDA is probably responsible for the immunosuppressive activities previously observed for the SAF. The strong affinity of SDA for proteins and thiocompounds may account for the apparent high MW previously reported for SAF.

Aldehydes↗

[Determination of two major metabolites of A-OT-Fu in mice serum by 4th order derivative UV spectrophotometry].

A 4th order derivative UV spectrophotometric method was developed for the determination of two major metabolites (TFu and 5-Fu) of A-OT-Fu in mice serum following per os of 290 mg/kg of the drug. TFu and 5-Fu were extracted from mice serum by ethyl acetate. The amplitudes of 252 and 289 nm for axle are linear with concentration of TFu and 5-Fu within the range of 5-50 micrograms/ml. Under the given experimental condition the recovery of TFu and 5-Fu were over 95%. The maximum concentration of TFu and 5-Fu appeared at 1 h and 2 h respectively after administration.

Animals↗

[Animal model and multiple trait BLUP applied in poultry genetic evaluation].

Best linear unbiased prediction (BLUP) is a powerful method to estimate genetic values of animals, and is widely applied in many animal species but poultry. Beijing White Leghorn nested data in 1986-1987 with 777 individuals were analysed by animal model and multiple trait BLUP. Two traits (40-week egg production and 36-week egg weight) and two fixed effects (house-pen effect and hatching batch effect) were considered. The way to calculate a large set of mixed model equations in micro-computer was studied. Only non-zero elements of the coefficient matrix of MME were stored on a disk. The iteration process was reduced by block iteration. It also simplified the multiple trait BLUP method, the dimension of equations is only 1/q (q is the number of traits) of regular method. So it saved a lot of compute time and cost, and BLUP became applicable in poultry. The significance of using BLUP in poultry are: (1) Eliminating some fixed effects; (2) Reducing the estimation error for unbalanced data; (3) Estimating breeding values of progeny, so we can shorten the generation interval; (4) It can estimate breeding values of individuals without records from the relatives' information; (5) The breeding value of sires and dams can be estimated from their progeny records, used for family selection; (6) Due to genetic and environmental correlation between traits and all relatives information were considered, it can increase the selection accuracy.

Animals↗

The purine nucleotide cycle and ammoniagenesis in rat kidney tubules.

The contribution of the purine nucleotide cycle to renal ammoniagenesis was examined in cortical tubule suspensions prepared from acidotic rats and incubated with [alpha-15N]glutamine, [15N]glutamate, or [15N]aspartate. Labeling of ammonia and adenine nucleotides was determined after enzymatic transformations designed to circumvent the technical problem that 15NH3 and H2O have the same nominal mass. Labeling of the adenine nucleotide was undetectable (less than 10%) even after 1 h of incubation. From the measured concentrations of adenine nucleotides and ammonia and the labeling of the ammonia, the flux through the purine nucleotide cycle was calculated to account for less than 1% of the deamination of alpha-amino groups from all three substrates. The glutamate dehydrogenase reaction is therefore the likely pathway for deamination. The rate of 15NH3 production from [alpha-15N]glutamine was two or three times greater than from added [15N]glutamate, indicating a preference for intracellularly generated glutamate. 15NH3 production from added [15N]aspartate was similar to and perhaps slightly greater than that from added [15N]glutamate.

Adenine Nucleotides↗

Two tissue-specific isozymes of creatine kinase have closely matched amino acid sequences.

Creatine kinase activity is associated with different isozyme species. We have examined two of these: the cytoplasmic brain (B) isozyme that is expressed in many tissues and is reported to be induced by estrogen and the developmentally regulated cytoplasmic muscle (M) isozyme that is found predominantly in differentiated muscle tissue. Recently, we cloned and sequenced the cDNA for the M isoenzyme of rabbit creatine kinase. We now report the isolation of B-isozyme cDNAs and the deduced primary structure of the polypeptide. The translated cDNA nucleotide sequence was cross-checked by fast-atom bombardment/mass spectrometry of tryptic fragments from the protein. The sequence is exactly colinear with the rabbit M isozyme and the two isozymes have 80% nucleotide and amino acid sequence identity. There are blocks of 36 and 41 amino acids where the amino acid sequence is conserved exactly. The colinearity of the two sequences and the extent of their identity makes it unlikely that either isozyme has unique polypeptide domains that account for specialized functions. The rationale for the existence of these creatine kinase isozymes, with distinct biological features, evidently is at the level of regulation of individual isozyme expression.

Amino Acid Sequence↗

Structural elucidation of a unique macrocyclic membrane lipid from a new, extremely thermophilic, deep-sea hydrothermal vent archaebacterium, Methanococcus jannaschii.

The membrane lipid of a new deep-sea hydrothermal vent methanogen, Methanococcus jannaschii, was isolated, purified, and structurally characterized. The total lipid extract, amounting to 32.2 micrograms/mg, dry cell weight, was fractionated on silica gel into a neutral lipid and polar lipid fraction. The neutral lipid fraction consisted of a series of isoprenoid hydrocarbons and free (nonphospholipid) alkylglycerol ethers. The polar phospholipid and glycolipid fraction (8.44 micrograms/mg, dry cell weight) was hydrolyzed with methanolic HCl, and the resulting alkylglycerol ethers were analyzed by a combination of chemical and spectroscopic techniques. The hydrolyzed polar lipid was primarily (95%) a unique, macrocyclic glycerol diether, heretofore unknown. High-field (250 MHz) proton nuclear magnetic resonance and infrared spectra of this novel macrocyclic compound are nearly identical and overlapping those of the known bis-(phytanyl)glycerol diether and bis-(diphytanyl)diglycerol tetraether. A field desorption mass spectrum revealed a molecular weight of 650 for the macrocyclic glycerol diether, 2 mass units less than that of bis-(phytanyl)glycerol diether. Degradation of the macrocyclic ether with boron tribromide resulted in diphytanyl dibromide, and further reaction of this dibromide with lithium aluminium hydride resulted in diphytane as determined by gas chromatography-mass spectrometry. The significance of the predominance of this structure in M. jannaschii is discussed. A survey of selected methanogenic Archaebacteria, including three thermophiles, failed to indicate the presence of the macrocyclic glycerol diether in any other microorganism, including two species of order Methanococcales, one species of Methanobacteriales, and three strains belonging to the order Methanomicrobiales.

Chemical Phenomena↗

Rabbit muscle creatine phosphokinase. CDNA cloning, primary structure and detection of human homologues.

A cDNA library was constructed from rabbit muscle poly(A) RNA. Limited amino acid sequence information was obtained on rabbit muscle creatine phosphokinase and this was the basis for design and synthesis of two oligonucleotide probes complementary to a creatine kinase cDNA sequence which encodes a pentapeptide. Colony hybridizations with the probes and subsequent steps led to isolation of two clones, whose cDNA segments partially overlap and which together encode the entire protein. The primary structure was established from the sequence of two cDNA clones and from independently determined sequences of scattered portions of the polypeptide. The reactive cysteine has been located to position 282 within the 380 amino acid polypeptide. The rabbit cDNA hybridizes to digests of human chromosomal DNA. This reveals a restriction fragment length polymorphism associated with the human homologue(s) which hybridizes to the rabbit cDNA.

Amino Acid Sequence↗