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Biomedical subjects

J W Webb

Publications and source records attributed to J W Webb.

15 recordsLinked to original sources
PubMed1995

Rapid mass spectrometric peptide sequencing and mass matching for characterization of human melanoma proteins isolated by two-dimensional PAGE.

We report a general mass spectrometric approach for the rapid identification and characterization of proteins isolated by preparative two-dimensional polyacrylamide gel electrophoresis. This method possesses the inherent power to detect and structurally characterize covalent modifications. Absolute sensitivities of matrix-assisted laser desorption ionization and high-energy collision-induced dissociation tandem mass spectrometry are exploited to determine the mass and sequence of subpicomole sample quantities of tryptic peptides. These data permit mass matching and sequence homology searching of computerized peptide mass and protein sequence data bases for known proteins and design of oligonucleotide probes for cloning unknown proteins. We have identified 11 proteins in lysates of human A375 melanoma cells, including: alpha-enolase, cytokeratin, stathmin, protein disulfide isomerase, tropomyosin, Cu/Zn superoxide dismutase, nucleoside diphosphate kinase A, galaptin, and triosephosphate isomerase. We have characterized several posttranslational modifications and chemical modifications that may result from electrophoresis or subsequent sample processing steps. Detection of comigrating and covalently modified proteins illustrates the necessity of peptide sequencing and the advantages of tandem mass spectrometry to reliably and unambiguously establish the identity of each protein. This technology paves the way for studies of cell-type dependent gene expression and studies of large suites of cellular proteins with unprecedented speed and rigor to provide information complementary to the ongoing Human Genome Project.

Amino Acid Sequence↗
PubMed1989

Structure and heterogeneity of the oligosaccharides from the lipopolysaccharides of a pyocin-resistant Neisseria gonorrhoeae.

The compositions and partial structures of the oligosaccharides from the lipopolysaccharides (LPS) of a pyocin-resistant Neisseria gonorrhoeae (strain JW31R) have been determined by liquid secondary ion mass spectrometry (LSIMS), tandem mass spectrometry, and methylation analysis. Four major structures were identified with Mr 2123, 2000, 1961, and 1838, as well as seven species of lower abundance of Mr 1758-1272. The largest of the major oligosaccharides (Mr, 2122) consists of 3-deoxymanno-2-ketooctulosonic acid (KDO)-Hep2GalNAcGlcNAcGal4Glc2 (Hep, heptose) and phosphoethanolamine (PEA). The smaller oligosaccharides are truncated versions of this larger oligosaccharide. The oligosaccharides consist of a common triantennary structure containing KDO at the reducing terminus attached to a heptose disaccharide. A hexose (Hex)2-3 branch is attached to the heptose linked directly to KDO and a GalNAc-Hex3, GlcNAc, and PEA are separately attached to the second heptose. These oligosaccharides are the first structures to be determined for a gonococcal LPS and should further our understanding of the structural and antigenic diversity of these glycolipids.

Bacteriocins↗
PubMed1988

Structural characterization of intact, branched oligosaccharides by high performance liquid chromatography and liquid secondary ion mass spectrometry.

We report results of a mass-spectrometric-based strategy for determining the detailed structural features of N-linked oligosaccharides from glycoproteins. The method was used to characterize a series of intact, high mannose oligosaccharides isolated from human immunoglobulin M (IgM). The IgM was purified from a patient with Waldenstrom's macroglobulinemia. The strategy included releasing the oligosaccharides by digestion of the purified glycoprotein with endoglycosidase H, separating the released oligosaccharides by high resolution gel filtration, and derivatizing the resulting reducing termini with the uv-absorbing moiety, ethyl p-aminobenzoate. This particular derivative facilitates HPLC detection and provides centers for protonation and deprotonation enhancing liquid secondary ion mass spectra. Positive and negative ion spectra contained molecular species of similar abundance. However, fragment ion peaks yielding sequence information were significantly more prominent in the negative ion mass spectra. Furthermore, it was obvious that the fragmentation patterns differed substantially for linear and branched oligomers. For linear oligosaccharides, a smooth envelope of fragment ions was observed; from low to high mass there was an ordered decrease in ion abundance from both the reducing and nonreducing termini. This pattern of fragment ions was not observed for branched oligosaccharides since in these cases fragments at certain masses could not arise by single bond cleavages. Therefore, these fragments were either significantly reduced in abundance or absent as compared with identical fragments formed from linear molecules. Importantly, 200 pmol of an oligosaccharide could be derivatized, separated, and detected by mass spectrometry, allowing identification of previously unreported minor components of the IgM oligosaccharides. Therefore, this experimental strategy is particularly useful for the purification and detailed structural characterization of low abundance oligosaccharides isolated from heterogeneous biological samples.

Carbohydrate Conformation↗
PubMed1986

Harvey A.K. Whitney lecture. The art of growing professionally.

The paths for professional development are described using past recipients of the Harvey A.K. Whitney Lecture Award as examples. Past recipients were able to improve their performance by accepting the help of others, including their families. Most of these individuals were guided by mentors who taught them how to think and supported their character. Their growth was enhanced through memberships in professional associations. As members of regional and national organizations, they were able to observe pharmacy practice outside their own communities. An attempt to grow professionally is in effect a search for excellence. One way that past recipients achieved excellence was by participating in continuing education. One of the most important factors contributing to their professional growth was their relationships with other people. Hospital pharmacists have been influenced by such persons as Harvey A.K. Whitney, Donald Francke, Donald Brodie, and Evlyn Gray Scott. Past recipients of the Harvey A.K. Whitney Lecture Award have exemplified professional growth.

Pharmacists↗
PubMed1984

Metabolic profiles of rats of different ages.

Urine samples from 6- to 31-month-old male Fischer F344 rats were analyzed using a high performance liquid chromatograph and a unique computer-based data analysis and quality control system in order to discover substances that could be used as markers in the aging process. Metabolic profiles of the organic acids from these urines yielded 42 peaks whose areas could be measured reliably. Of the 42 peaks, 10 were found by analysis of variance to vary significantly (p less than 0.05) with age. Rats from the four ages could also be distinguished using a multivariate statistic (discriminant analysis).

Aging↗
PubMed1982

Comparison of isolation methods of urinary organic acids by high-performance liquid chromatography.

Four methods for extracting organic acids from human urine prior to analysis by high-performance liquid chromatography (HPLC) were compared. The methods were manual solvent extraction with ethyl acetate and diethyl ether, continuous solvent extraction, anion exchange with pyridinium acetate as the eluting solvent and anion exchange with hydrochloric acid as the eluting solvent. All four methods produced samples that could be analyzed by reversed-phase HPLC, but the continuous solvent extraction and anion exchange with pyridinium acetate methods gave the best reproducibilities (approximately 6% relative standard deviations). Pretreatment of the urine with barium hydroxide and hydroxylamine hydrochloride prior to anion exchange did not markedly alter the HPLC profiles.

Adult↗
PubMed1976

Compatibility of penicillin and ascorbic acid injection.

The stability of Potassium Penicillin G, USP, when mixed with Ascorbic Acid Injection, USP, in 5% Dextrose Injection, USP, was studied. The change in concentration over an eight-hour period of potassium penicillin G in the admixture was determined by the hydroxylamine colorimetric assay method and the microbiological assay method. The stability of penicillin was not adversely affected by the presence of sodium ascorbate. Reports of incompatibilities between penicillin and ascorbic acid are a function of pH rather than a characteristic of the ascorbate ion. Articles reporting studies involving ascorbic acid should specify whether the work refers to the use of ascorbic acid or Ascorbic Acid Injection, USP. Confusion in the literature could be reduced by changing the official title from Ascorbic Acid Injection to Sodium Ascorbate Injection.

Ascorbic Acid↗