Identification of In-Chain-Functionalized Compounds and Methyl-Branched Alkanes in Cuticular Waxes of Triticum aestivum cv. Bethlehem

In this work, cuticular waxes from flag leaf blades and peduncles of Triticum aestivum cv. Bethlehem were investigated in search for novel wax compounds. Seven wax compound classes were detected that had previously not been reported, and their structures were elucidated using gas chromatography-mass spectrometry of various derivatives. Six of the classes were identified as series of homologs differing by two methylene units, while the seventh was a homologous series with homologs with single methylene unit differences. In the waxes of flag leaf blades, secondary alcohols (predominantly C₂₇ and C₃₃), primary/secondary diols (predominantly C₂₈) and esters of primary/secondary diols (predominantly C₅₀, combining C₂₈ diol with C₂₂ acid) were found, all sharing similar secondary hydroxyl group positions at and around C-12 or ω-12. 7- and 8-hydroxy-2-alkanol esters (predominantly C₃₅), 7- and 8-oxo-2-alkanol esters (predominantly C₃₅), and 4-alkylbutan-4-olides (predominantly C₂₈) were found both in flag leaf and peduncle wax mixtures. Finally, a series of even- and odd-numbered alkane homologs was identified in both leaf and peduncle waxes, with an internal methyl branch preferentially on C-11 and C-13 of homologs with even total carbon number and on C-12 of odd-numbered homologs. Biosynthetic pathways are suggested for all compounds, based on common structural features and matching chain length profiles with other wheat wax compound classes.     

Conformation of poly(L-lysine) homologs in solution

The effect of the number of methylene groups in the side chains on the conformation of polypeptides is assessed for three poly(L -lysine) homologs with R = –(CH₂)_nNH₂. Circular dichroism studies show a pH-induced helix–coil transition in 0.05 M KCl with midpoints at 9.6, 9.0, and 8.7 for n = 5, 6, and 7, respectively, as compared with 10.1 for (Lys)_x (n = 4). Homologs with n = 6 and 7 could be partially helical even when the side groups are fully charged (with n = 7, the compound is highly aggregated above pH 9.1). Thus, the longer the number of methylene groups the more stable is the helical conformation of these homologs. Potentiometric titration of the n = 5 homolog gives a ΔG° of ?310 cal/mol (residue) for the uncharged coil-to-helix transition at 25°C. The corresponding ΔH° and ΔS° are ?1740 cal/mol (residue) and ?4.8 e.u./mol (residue). Unlike (Lys)_x, the uncharged helix-to-β transition is slow and incomplete even after heating at 80°C for 1 hr. Addition of methanol enhances the helical formation in neutral solution with midpoints at 72, 52, and 27% methanol (v/v) for n = 5, 6, and 7, respectively [cf. 88% for (Lys)_x]. Addition of sodium dodecyl sulfate induces a coil-to-helix transition for all three homologs in contrast with the β form of (Lys)_x under similar conditions.     

THE ORIGIN OF MORPHOLOGICAL CHARACTERS AND THE BIOLOGICAL BASIS OF HOMOLOGY

A homolog is a part of the phenotype that is homologous to equivalent parts in other species. A biological homology concept is expected to explain three properties of homologs: 1) the conservation of those features that are used to define a homolog, 2) the individualization of the homolog with regard to the rest of the body, and 3) the uniqueness of homologs, i.e., their specificity for monophyletic groups. The main obstacle to describing a mechanistic basis for homology is the variability of the developmental pathways of undoubtedly homologous characters. However, not all aspects of the developmental pathway are of equal importance. The only organizational features of the developmental system that matter are those that have been historically acquired and cause developmental constraints on the further evolutionary modification of the characters. Two main factors contribute to historically acquired developmental constraints: generative rules of pattern formation and ontogenetic networks. In particular, hierarchical and cyclical inductive networks have the required properties to explain homology. How common such networks are is an open empirical question. The development and variation of pectoral fin hooks in blenniid fishes is presented as a model for the study of a simple ontogenetic network.     

Bacteriochlorophyll c monomers,dimers, and higher aggregates in dichloromethane,chloroform, and carbon tetrachloride

Bacteriochlorophyll c in vivo is a mixture of at least 5 homologs, all of which form aggregates in CH₂Cl₂, CHCl₃ and CCl₄. Three homologs exist mainly in the 2-R-(1-hydroxyethyl) configuration, whereas the other two homologs, 4-isobutyl-5-ethyl and 4-isobutyl-5-methyl farnesyl bacteriochlorophyll c, exist mainly in the 2-S-(1-hydroxyethyl) configuration (Smith KM, Craig GW, Kehres LA and Pfennig N (1983) J. Chromatograph. 281: 209–223). In CCl₄ the S-homologs form an aggregate of 2–3 molecules whose absorption (747 nm maximum) and circular dichroism spectra resemble those of the chlorosome. In CH₂Cl₂, CHCl₃ and CCl₄ the 4-n-propyl homolog (R-configuration) forms dimers absorbing at ca. 680 nm and higher aggregates absorbing at 705–710 nm. In CCl₄ the dimerization constant is approx. 10 µM^–1 (1000 times that for chlorophyll a). The difference between the types of aggregates formed by the 4-n-propyl and 4-isobutyl homologs is attributed to the difference between the R- and S-configurations of the 2-(1-hydroxyethyl) groups in each chlorophyll.Abbreviations BChl bacteriochlorophyll - CD circular dichroism - Chl chlorophyll - DNS data not shown - EEF 4-ethyl-5-ethyl farnesyl - iBM/EF 4-isobutyl-5-methyl/ethyl farnesyl - MEF 4-methyl-5-ethyl farnesyl - PEP 4-n-propyl-5-ethyl farnesyl     

Effect of chain length and concentration of anionic surfactants on the conformational transitions of poly(L-ornithine) and poly(L-lysine) in aqueous solution

The conformation of poly(L-ornithine) (PLO) and poly(L-lysine) (PLL) in solutions of sodium alkyl sulfates, CH₃(CH₂)_nSO₄Na with n = 7, 9, 11, 13 and 15 was studied by circular dichroism. PLO adopts a helical conformation in all 5 homologs and PLL a β-form in only 4 of the homologs. With octyl sulfate PLL has a helical conformation instead. These conformations were observed in solution of surfactants both below and above the critical micelle concentration.     

Synthesis,physicochemical and biological evaluation of tacrine derivative labeled with technetium-99m and gallium-68 as a prospective diagnostic tool for early diagnosis of Alzheimer’s disease

Design, physicochemical and biological studies of novel radioconjugates for the early diagnosis of Alzheimer's disease, based on the newly synthesized tacrine derivatives were performed. Novel tacrine analogues were labeled with technetium-99m and gallium-68. For all obtained radioconjugates ([^99mTc]Tc-Hynic-(tricine)₂NH(CH₂)_ntacrine and [⁶⁸Ga]Ga-DOTA-NH(CH₂)₉tacrine, where n = 2–9 denotes the number of methylene groups CH₂) the studies of physicochemical properties (lipophilicity, stability in the presence of an excess of standard amino acids cysteine or histidine, human serum and in cerebrospinal fluid) were performed. For two selected radioconjugates [^99mTc]Tc-Hynic-(tricine)₂NH(CH₂)₉Tac and [⁶⁸Ga]Ga-DOTA-NH(CH₂)₉tacrine (characterized with the highest lipophilicity values) the biological tests (inhibition of cholinesterases action, molecular docking and biodistribution studies) have been performed. All novel radioconjugates showed high stability in biological solutions used. Both selected radioconjugates proved to be good inhibitors of cholinesterases and be able to cross the blood-brain barrier. Radioconjugates [^99mTc]Tc-Hynic-(tricine)₂NH(CH₂)₉tacrine and [⁶⁸Ga]Ga-DOTA-NH(CH₂)₉tacrine fulfil the conditions for application in nuclear medicine. Radiopharmaceutical [⁶⁸Ga]Ga-DOTA-NH(CH₂)₉tacrine, due to increased accuracy and improved sensitivity in PET imaging, may be better potential diagnostic tool for early diagnosis of Alzheimer’s disease.     

A DFT study of the effect of K and SiO2 on syngas conversion to methane and methanol over an Mo6P3 cluster

Synthesis gas (CO+H₂) conversion to CH₄ and CH₃OH over an Mo₆P₃ cluster, an Mo₆P₃–Si₃O₉ and a K–Mo₆P₃–Si₃O₉ cluster has been studied using density functional theory (DFT). The study focused on the reaction between the intermediate species CH₂OH_ad+H_ad, comparing methanol formation to C–O bond scission that yields CH_2.ad+H₂O_ad species. The activation energies of both the reactions decreased on the Mo₆P₃–Si₃O₉ and the K–Mo₆P₃–Si₃O₉ clusters compared to the Mo₆P₃ cluster. However, on the K–Mo₆P₃–Si₃O₉ cluster, the activation energy for methanol formation (12.1 kcal/mol) was higher than the C–O bond-breaking activation energy (9.9 kcal/mol). Although the DFT study predicted preferential formation of CH₄ versus CH₃OH on all the Mo₆P₃ clusters, the study also predicted an increased formation of CH₃OH with the addition of K and experimental measurements are in agreement with this prediction.     

The cytokinin activities of 6-α-alkylbenzyloxypurines

The homologous series of 6-α-n-alkylbenzyloxypurines with alkyl groups (—(CH₂)_nCH₃) from n = 0 to n = 11 have been prepared and examined in three tests for cytokinin activity. The parent benzyloxypurine was also included in the tests. Substituting methyl (n = 0) into the methylene group of 6-benzyloxypurine removed activity almost completely; thereafter, increasing the size of the alkyl group (n = 1–5) gave compounds which were active in all tests, the butyl (n = 3) and pentyl (n = 4) derivatives being more active than 6-benzyloxypurine itself. Activity then fell as the series was ascended; the higher homologues (n = 8–11) being completely inactive. The results are discussed in relation to steric and other considerations.     

Diverse communities of Bacteria and Archaea flourished in Palaeoarchaean (3.5–3.3 Ga) microbial mats

Limited taxonomic classification is possible for Archaean microbial mats and this is a fundamental limitation in constraining early ecosystems. Applying Fourier transform infrared spectroscopy (FTIR), a powerful tool for identifying vibrational motions attributable to specific functional groups, we characterized fossilized biopolymers in 3.5–3.3 Ga microbial mats from the Barberton greenstone belt (South Africa). Microbial mats from four Palaeoarchaean horizons exhibit significant differences in taxonomically informative aliphatic contents, despite high aromaticity. This reflects precursor biological heterogeneity since all horizons show equally exceptional preservation and underwent similar grades of metamorphism. Low methylene to end-methyl (CH₂/CH₃) absorbance ratios in mats from the 3.472 Ga Middle Marker horizon signify short, highly branched n-alkanes interpreted as isoprenoid chains forming archaeal membranes. Mats from the 3.45 Ga Hooggenoeg Chert H5c, 3.334 Ga Footbridge Chert, and 3.33 Ga Josefsdal Chert exhibit higher CH₂/CH₃ ratios suggesting mostly longer, unbranched fatty acids from bacterial lipid precursors. Absorbance ratios of end-methyl to methylene (CH₃/CH₂) in Hooggenoeg, Josefsdal and Footbridge mats yield a range of values (0.20–0.80) suggesting mixed bacterial and archaeal architect communities based on comparison with modern examples. Higher (0.78–1.25) CH₃/CH₂ ratios in the Middle Marker mats identify Archaea. This exceptional preservation reflects early, rapid silicification preventing the alteration of biogeochemical signals inherited from biomass. Since silicification commenced during the lifetime of the microbial mat, FTIR signals estimate the affinities of the architect community and may be used in the reconstruction of Archaean ecosystems. Together, these results show that Bacteria and Archaea flourished together in Earth's earliest ecosystems.     

Structural Specificity of Polyamines in Modulating the Binding of Estrogen Receptor to Potential Z-DNA Forming Sequences

Abstract

Estrogen receptor (ER) is a gene-regulatory protein that mediates the action of estradiol. In order to examine the role of conformational dynamics of DNA in estrogenic regulation of gene expression, we studied the binding of ER to poly(dA-dC).poly(dG-dT) which undergoes transition to a left-handed Z-DNA form. This type of dinucleotide repeats are widely distributed in mammalian genome and are present in estrogen response elements. Binding affinity of ER for the polynucleotide was assessed by its ability to release ER bound to DNA-cellulose. ER binding by poly(dA-dC).poly(dG-dT) was enhanced in the presence of an endogenous polyamine, spermidine, H₂N(CH₂)₄NH(CH₂)₃NH₂. The concentration of spermidine required for facilitating 50% elution of ER (EC₅₀) was 75 μM. This EC₅₀ increased to 500 μM for a spermidine homolog, H₂N(CH₂)₈NH(CH₂)₃NH₂, demonstrating polyamine structural specificity. Spectroscopic measurements showed that the presence of 100 – 200 μM spermidine initiated changes in the conformation of the polynucleotide indicative of Z-DNA form, but a major alteration to Z-DNA spectrum occurred only at 300 μM concentration. These data suggest that ER favors DNA sequences poised for Z-DNA transition. The efficacy of spermidine homologs in facilitating ER-DNA interaction may be important in predicting their efficiency to replace cellular functions of spermidine.     

Analysis of homologous derivatives of cephalothin by multivariate methods for clinical efficacy

This work presents the analysis of pharmacological properties of a homologous set of cephalothin derivatives formed after inserting an aliphatic ester substituent having from one to ten carbon atoms (ie. -CH3 or -CH2CH3) in place of the former carboxyl group (-C(O)OH). These compounds were shown to have significant correlations and associations in their properties after analysis by pattern recognition methods including cluster analysis, detrended correspondence analysis, and K-means cluster analysis. Formula weight of all derivatives is directly correlated and increases with molar volume, parachor, and molar refractivity. Index of refraction decreases as formula weight of derivatives increases. Polar surface area of all derivatives remains constant at 102.02 A2 as formula weight increases. Partitioning between 1-octanol/water values of Log P increases as the length of the aliphatic ester group increases. The number of nitrogens, oxygens, -NH and -OH groups, remains constant for all derivatives remains the same at 2, 6, and 1, respectively. Homologs 1 to 7 (based on number carbon atoms of ester group) show zero violations of the Rule of 5, which indicates effective drug bioavailabilty. Values of polar surface area indicate that more than 25% of any derivative present in the intestinal system would be absorbed. The ethyl and propyl derivative of cephalothin have Log P values indicating efficient permeation of the central nervous system. Detrended correspondence analysis and K-means cluster analysis showed associations and interrelationships among these derivatives that will be clinically useful for the treatment of bacterial infections.     

An Intertwined Evolutionary History of Methanogenic Archaea and Sulfate Reduction

Hydrogenotrophic methanogenesis and dissimilatory sulfate reduction, two of the oldest energy conserving respiratory systems on Earth, apparently could not have evolved in the same host, as sulfite, an intermediate of sulfate reduction, inhibits methanogenesis. However, certain methanogenic archaea metabolize sulfite employing a deazaflavin cofactor (F₄₂₀)-dependent sulfite reductase (Fsr) where N- and C-terminal halves (Fsr-N and Fsr-C) are homologs of F₄₂₀H₂ dehydrogenase and dissimilatory sulfite reductase (Dsr), respectively. From genome analysis we found that Fsr was likely assembled from freestanding Fsr-N homologs and Dsr-like proteins (Dsr-LP), both being abundant in methanogens. Dsr-LPs fell into two groups defined by following sequence features: Group I (simplest), carrying a coupled siroheme-[Fe₄-S₄] cluster and sulfite-binding Arg/Lys residues; Group III (most complex), with group I features, a Dsr-type peripheral [Fe₄-S₄] cluster and an additional [Fe₄-S₄] cluster. Group II Dsr-LPs with group I features and a Dsr-type peripheral [Fe₄-S₄] cluster were proposed as evolutionary intermediates. Group III is the precursor of Fsr-C. The freestanding Fsr-N homologs serve as F₄₂₀H₂ dehydrogenase unit of a putative novel glutamate synthase, previously described membrane-bound electron transport system in methanogens and of assimilatory type sulfite reductases in certain haloarchaea. Among archaea, only methanogens carried Dsr-LPs. They also possessed homologs of sulfate activation and reduction enzymes. This suggested a shared evolutionary history for methanogenesis and sulfate reduction, and Dsr-LPs could have been the source of the oldest (3.47-Gyr ago) biologically produced sulfide deposit.     

New ways of determining structural groups in brown coals and their bioconversion products by FT IR spectroscopy

New methods of determining the structural groups —COOH and —CH₂— have been developed. The investigation of carboxyl groups is possible both after derivatization with p-fluorophenacylbromide and by quantitative interpretation of the Fourier transform infrared (FT IR) spectra. There exists a linear relationship between the results of these two methods that is generally valid for the analysis of all brown coal components. The maximum extinction coefficient of the symmetric stretching vibration band of the CH₂ groups has been determined using model substances. This allows quantification of this structural group directly from the FT IR spectrum. The results agree with the contents of methylene groups as determined by ¹³C-cross polarization–magic angle spinning–nuclear magnetic resonance (¹³C CPMAS NMR) spectroscopy. Using these methods, the COOH and CH₂ groups contained in brown coals of the North Rhine region and in their bioconversion products have been quantified. Received: 21 December 1999 / Received revision: 25 April 2000 / Accepted: 1 May 2000     

Conformational characteristics of polyglutamic acid esters: Persistence of orientational correlation along the side chain flanking the α-helical backbone

The side chain conformations of α-helical poly(L -glutamic acid) esters $ \rlap{--}[NHCH(CH_2 CH_2 COOR)CO\rlap{--}]_x $, carrying a homologous series of ester residues such as R = ? (CH₂)_n? with n = 1–3, have been studied in the lyotropic liquid crystalline state (chloroform 20 v/v%) by the deuterium nmr method. In order to study the surface chirality of the molecule, the phenyl groups situated at the terminal of the side chain have been deuterated. From the observed deuterium quadrupolar splittings, the average inclination θ_p of the para-axis of the phenyl group with respect to the α-helical backbone was elucidated. A distinct odd–even oscillation in the quantity such as 〈 cos² θ_p〉 was observed with the number of methylene units n. A rotational isomeric state analysis has indicated that the observed orientational correlation arises from the interdependence of the neighboring bond rotation along the side chain. Preference of the “extended” conformations is also enhanced by the mutual conformational exclusion of neighboring side chains.     

Single and double nucleophilic attack on cluster-bonded allenylidenes: Synthesis of phosphorus heterocycles

Further studies of the attack of bidentate tertiary phosphines, such as dppm and dppe, and the related diarsine dpam, on Ru₃(μ-H){μ₃-C₂CHR(OH)}(CO)₉ (R = H, Me) are described, together with the X-ray determined structures of [Ru₃(μ₃-C₂CH₂PPh₂CH₂CH₂PPh₂)(CO)₉]BF₄ (6) and Ru₃(μ-H){μ₃-C₂CHMePPh₂CHPPh₂}(CO)₉ (7) (containing diphospha heterocycles), Ru₃(μ-H){μ₃-CH₂(OH)C₂PPh₂CH₂PPh₂}(CO)₈ (8) (containing a phosphonium-alkynyl ligand also coordinated to a cluster Ru atom) and Ru₃(μ-H)(μ₃-CCCHAsPh₂CH₂AsPh₂)(CO)₉ (9) (containing an arsino-allenylidene ligand). These complexes are formed by nucleophilic attack of the Group 15 ligand on an alkynyl or allenylidene ligand on the cluster.     

Effect of Temperature on Carbon and Electron Flow and on the Archaeal Community in Methanogenic Rice Field Soil

Temperature is an important factor controlling CH₄ production in anoxic rice soils. Soil slurries, prepared from Italian rice field soil, were incubated anaerobically in the dark at six temperatures of between 10 to 37°C or in a temperature gradient block covering the same temperature range at intervals of 1°C. Methane production reached quasi-steady state after 60 to 90 days. Steady-state CH₄ production rates increased with temperature, with an apparent activation energy of 61 kJ mol⁻¹. Steady-state partial pressures of the methanogenic precursor H₂ also increased with increasing temperature from <0.5 to 3.5 Pa, so that the Gibbs free energy change of H₂ plus CO₂-dependent methanogenesis was kept at −20 to −25 kJ mol of CH₄⁻¹ over the whole temperature range. Steady-state concentrations of the methanogenic precursor acetate, on the other hand, increased with decreasing temperature from <5 to 50 μM. Simultaneously, the relative contribution of H₂ as methanogenic precursor decreased, as determined by the conversion of radioactive bicarbonate to ¹⁴CH₄, so that the carbon and electron flow to CH₄ was increasingly dominated by acetate, indicating that psychrotolerant homoacetogenesis was important. The relative composition of the archaeal community was determined by terminal restriction fragment length polymorphism (T-RFLP) analysis of the 16S rRNA genes (16S rDNA). T-RFLP analysis differentiated the archaeal Methanobacteriaceae, Methanomicrobiaceae, Methanosaetaceae, Methanosarcinaceae, and Rice clusters I, III, IV, V, and VI, which were all present in the rice field soil incubated at different temperatures. The 16S rRNA genes of Rice cluster I and Methanosaetaceae were the most frequent methanogenic groups. The relative abundance of Rice cluster I decreased with temperature. The substrates used by this microbial cluster, and thus its function in the microbial community, are unknown. The relative abundance of acetoclastic methanogens, on the other hand, was consistent with their physiology and the acetate concentrations observed at the different temperatures, i.e., the high-acetate-requiring Methanosarcinaceae decreased and the more modest Methanosaetaceae increased with increasing temperature. Our results demonstrate that temperature not only affected the activity but also changed the structure and the function (carbon and electron flow) of a complex methanogenic system.     

Electrogenerated chemiluminescence of the platinum (II) octaethylporphyrin/tri-n-propylamine system

The electrogenerated chemiluminescence (ECL) of platinum (II) octaethyl-porphyrin (PtOEP) in acetonitrile:methylene chloride (CH₃CN:CH₂Cl₂, 50:50 v/v) and CH₂Cl₂ is reported. ECL was generated upon sweep to positive potentials using tri-n-propylamine (TPrA) as an oxidative-reductive coreactant. ECL efficiencies (?_ecl) of 0.18 in CH₃CN:CH₂Cl₂ (50:50 v/v) and 3.90 in methylene chloride were obtained using Ru(bpy)₃(PF₆)₂ (bpy = 2,2′-bipyridine) as a relative standard (?_ecl = 1). The ECL intensity peaks at a potential corresponding to oxidation of PtOEP and TPrA, and ECL emission spectra are nearly identical to photoluminescence emission spectra, indicating that emission is from the PtOEP triplet state.     

Identification, Genomic Organization, and Analysis of the Group III Capsular Polysaccharide Genes kpsD, kpsM, kpsT, and kpsE from an Extraintestinal Isolate of Escherichia coli (CP9, O4/K54/H5)

Group III capsular polysaccharides (e.g., K54) of extraintestinal isolates of Escherichia coli, similar to group II capsules (e.g., K1), are important virulence traits that confer resistance to selected host defense components in vitro and potentiate systemic infection in vivo. The genomic organization of group II capsule gene clusters has been established as a serotype-specific region 2 flanked by regions 1 and 3, which contain transport genes that are highly homologous between serotypes. In contrast, the organization of group III capsule gene clusters is not well understood. However, they are defined in part by an absence of genes with significant nucleotide homology to group II capsule transport genes in regions 1 and 3. Evaluation of isogenic, TnphoA-generated, group III capsule-minus derivatives of a clinical blood isolate (CP9, O4/K54/H5) has led to the identification of homologs of the group II capsule transport genes kpsDMTE. These genes and their surrounding regions were sequenced and analyzed. The genomic organization of these genes is distinctly different from that of their group II counterparts. Although kps_K54DMTE are significantly divergent from their group II homologs at both the DNA and protein levels phoA fusions and computer-assisted analyses suggest that their structures and functions are similar. The putative proteins Kps_K54M and Kps_K54T appear to be the integral membrane component and the peripheral ATP-binding component of the ABC-2 transporter family, respectively. The putative Kps_K54E possesses features similar to those of the membrane fusion protein family that facilitates the passage of large molecules across the periplasm. At one boundary of the capsule gene cluster, a truncated kpsM (kpsM_truncated) and its 5′ noncoding regulatory sequence were identified. In contrast to the complete kps_K54M, this region was highly homologous to the group II kpsM. Fifty-three base pairs 3′ from the end of kpsM_truncated was a sequence 75% homologous to the 39-bp inverted repeat in the IS110 insertion element from Streptomyces coelicolor. Southern analysis established that two copies of this element are present in CP9. These findings are consistent with the hypothesis that CP9 previously possessed group II capsule genes and acquired group III capsule genes via IS110-mediated horizontal transfer.     

Fowlpox Virus Encodes Nonessential Homologs of Cellular Alpha-SNAP, PC-1, and an Orphan Human Homolog of a Secreted Nematode Protein

The genome of fowlpox virus (FWPV), type species of the Avipoxviridae, is considerably rearranged compared with that of vaccinia virus (the prototypic poxvirus and type species of the Orthopoxviridae) and is 30% larger. It is likely that the genome of FWPV contains genes in addition to those found in vaccinia virus, probably involved with its replication and survival in the chicken. A 7,470-bp segment of the FWPV genome has five open reading frames (ORFs), two of which encode ankyrin repeat proteins, many examples of which have been found in poxviruses. The remaining ORFs encode homologs of cellular genes not reported in any other virus. ORF-2 encodes a homolog of the yeast Sec17p and mammalian SNAP proteins, crucial to vesicular transport in the exocytic pathway. ORF-3 encodes a homolog of an orphan human protein, R31240_2, encoded on 19p13.2. ORF-3 is also homologous to three proteins (YLS2, YMV6, and C07B5.5) from the free-living nematode Caenorhabditis elegans and to a 43-kDa antigen from the parasitic nematode Trichinella spiralis. ORF-5 encodes a homolog of the mammalian plasma cell antigen PC-1, a type II glycoprotein with exophosphodiesterase activity. The ORFs are present in the virulent precursor, HP1, of the sequenced attenuated virus (FP9) and are conserved in other strains of FWPV. They were shown, by deletion mutagenesis, to be nonessential to virus replication in tissue culture. RNA encoding the viral homolog of PC-1 is expressed strongly early and late in infection, but RNAs encoding the homologs of SNAP and R31240_2 are expressed weakly and late.     

Towards functional proteomics of minority component of honeybee royal jelly: The effect of post‐translational modifications on the antimicrobial activity of apalbumin2

This study illustrates multifunctionality of proteins of honeybee royal jelly (RJ) and how their neofunctionalization result from various PTMs of maternal proteins. Major proteins of RJ, designated as apalbumins belong to a protein family consisting of nine members with M_r of 49–87 kDa and they are accompanied by high number of minority homologs derived from maternal apalbumins. In spite of many data on diversity of apalbumins, the molecular study of their individual minority homologous is still missing. This work is a contribution to functional proteomics of second most abundant protein of RJ apalbumin2 (M_r 52.7 kDa). We have purified a minority protein from RJ; named as apalbumin2a, differ from apalbumin2 in M_r (48.6 kDa), in N‐terminal amino acids sequences – ENSPRN and in N‐linked glycans. Characterization of apalbumin2a by LC‐MALDI TOF/TOF MS revealed that it is a minority homolog of the major basic royal jelly protein, apalbumin2, carrying two fully occupied N‐glycosylation sites, one with high‐mannose structure, HexNAc2Hex9, and another carrying complex type antennary structures, HexNAc4Hex3 and HexNAc5Hex4. We have found that apalbumin2a inhibit growth of Paenibacillus larvae. The obtained data call attention to functional plasticity of RJ proteins with potential impact on functional proteomics in medicine.