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N G Anderson

Publications and source records attributed to N G Anderson.

At least 91 records · Page 5Linked to original sources
PubMed1982

Proteins of human milk. I. Identification of major components.

Traditionally, human milk proteins are identified largely by reference to bovine milk. Hence, to identify the major proteins in human milk, we subjected human and bovine milk, in parallel, to high-resolution two-dimensional electrophoresis. Isoelectric precipitation at pH 4.6 was our criterion for distinguishing whey proteins from those of the casein complex. The alpha- and beta-caseins were identified on the basis of relative abundance, relative molecular mass, and relative isoelectric points. Kappa casein was identified as a series of four spots, which disappear from bovine skim milk treated with rennin (chymosin; EC 3.4.23.4) during the clotting process. Para kappa-casein does not appear on the standard ISO-DALT pattern after treatment of bovine milk with rennin, but does appear in BASO-DALT pattern, indicating its high isoelectric point. No protein disappeared from ISO-DALT patterns of human milk after rennin treatment, and no new protein comparable to bovine para kappa-casein appeared in the BASO-DALT patterns; this suggests that kappa-casein is absent from human milk. The proteins identified in human milk patterns include the alpha and beta casein families, lactalbumin, albumin, transferrin, IgA, and lactoferrin. Numerous additional proteins seen in patterns for human milk remain to be identified.

Albumins↗
PubMed1982

Proteins of human urine. III. Identification and two-dimensional electrophoretic map positions of some major urinary proteins.

We mapped the proteins of human urine by high-resolution two-dimensional electrophoresis, utilizing the ISO-DALT system. Wide-range pH gradients and narrow-range acid gradients were both used in the first-dimension separations. The patterns revealed proteins ranging in relative molecular mass from 10 000 to 90 000. Proteins identified in the map included transferrin, albumin, hemopexin, alpha 2-HS glycoprotein, alpha 1-antitrypsin. Gc globulin, alpha 1-acid glycoprotein, Zn alpha 2-glycoprotein, retinol binding protein, beta 2-microglobulin, the immunoglobulin light chains, and MAUP (most acid urinary protein). The use and utility of internal-charge and molecular-mass standards are described. We used electrophoretic transfer of proteins to nitrocellulose sheets and subsequent detection by immunological methods to identify some proteins.

Animals↗
PubMed1982

Design and implementation of a prototype Human Protein Index.

This paper describes information-handling aspects of the TYCHO I analysis system (Clin, Chem. 27: 1807--1820, 1981), which analyzes two-dimensional electrophoresis gels, matches the individual protein spots with those in a reference pattern, and stores various information--including spot measurements, identifications, treatment profiles, set memberships, and comments--in a computerized database. This and additional information such as amino acid composition and cellular localization is then accessible from an interactive program that includes a pictorial user interface and presents much of the data in graphical form. Use of the TYCHO I system is illustrated by examples drawn from analyses of gel patterns from human leukocytes.

Blood Proteins↗
PubMed1982

Proteins of human urine. II. Identification by two-dimensional electrophoresis of a new candidate marker for prostatic cancer.

A protein series common to the urine and prostatic tissue of 16 of 17 patients with prostatic adenocarcinoma has been identified by high-resolution two-dimensional gel electrophoresis. These proteins, designated PCA-1, have a relative molecular mass in sodium dodecyl sulfate of about 40000. Analyses of urines from eight age-matched controls, seven patients with other ty pes of urogenital malignancies, two patients with benign prostatic hyperplasia, and five patients with malignancies not associated with the urogenital system failed to show PCA-1 in the patterns. These preliminary findings suggest that this protein should be systematically investigated as a candidate marker for prostatic adenocarcinoma in man.

Adenocarcinoma↗
PubMed1982

A two-dimensional electrophoretic analysis of the heat-shock-induced proteins of human cells.

Using two-dimensional electrophoresis, we have investigated the responses of human cells in culture to heat shock and to various chemical agents producing a similar effect. These treatments result in the induction of increased synthesis of several specific proteins. One (HShock:1, SDS-molecular mass about 65000) is increased by about 350-fold over the amount in untreated cells. Computer analysis of time-course studies indicates, however, that rates of synthesis of various proteins other than the classical "heat shock proteins" are affected, some of these alterations following time courses quite different from the main (HShock) inductions. The heat shock effect is thus much more complicated than previously realized. We purified the HShock:1 protein from heat-shocked human lymphoblastoid cells, and prepared a rabbit antiserum specific for HShock:1 on nitrocellulose two-dimensional gel transfers of total lymphoblastoid cell protein. A survey of mouse tissues shows high concentrations of an HShock:1-like protein in the testis, and human testes also appears to contain substantial (though lower) concentrations. These results are consistent with the hypothesis (derived from the tissue-culture studies) that the heat shock effect is a general response to the need for increased protein catabolism within the cell. Increased concentrations of HShock:1 are also observed in preparations of blood leukocytes collected from patients after surgery, indicating that some types of physiological trauma may induce the heat shock proteins in man. Using the antiHShock:1 antibody in an immunoassay, it will be possible to systematically examine HShock:1 concentrations in plasma and leukocytes, thereby opening up the possibility of a clinical test based for the first time upon an inducible aspect of cellular gene expression.

Animals↗
PubMed1981

Two-dimensional analysis of human lymphocyte proteins: I. An assay for lymphocyte effectors.

We describe an assay for lymphocyte effectors that is capable of establishing the existence of regulators of lymphocyte gene expression (including post-transcriptional control and protein processing) and has the ability to characterize the response at the molecular level. The hypothesis that circulating effectors substances excreted through the kidney can be actively present in human urine was tested with this assay. Thus, biologically active protein molecules in urine were detected at concentrations of less than 1 mg/L and over a wide range of dilutions. Activities were detected and quantitated by culturing human lymphocytes with human urinary proteins in the presence of [35S]methionine and subsequently analyzing the labeled lymphocyte proteins by two-dimensional gel electrophoresis. Thus, protein analysis by two-dimensional gels was used to indirectly detect changes produced in cultured lymphocytes after exposure to regulatory molecules. Proteins or sets of lymphocyte proteins appeared or disappeared after exposure to normal or pathological human urinary proteins. Normal human urinary proteins triggered the appearance of sets of proteins referred to by number as the "Urocon" proteins and suppressed the synthesis of protein sets referred to as "Urocof" proteins. In addition to the normal alterations described, urinary proteins from individuals with influenza or acute leukemia and after renal transplantation were capable of inducing unique alterations in lymphocyte patterns.

Blood Proteins↗
PubMed1981

Proteins of human semen. I. Two-dimensional mapping of human seminal fluid.

The proteins in human seminal plasma were mapped by high-resolution two-dimensional electrophoresis (ISO-DALT and BASO-DALT systems). When analyzed under dissociating conditions, samples from normal fertile males revealed a pattern of over 200 proteins, ranging in mass from 10 000 to 100 000 daltons. Comparison of the mapped proteins from these males and those who had undergone vasectomy allowed us to identify one series of glycoproteins as missing from the semen from vasectomized individuals. Glycoproteins isolated by affinity chromatography with use of concanavalin A were also mapped. Some of the protein spots were identified either by co-electrophoresis with purified proteins or by the electrophoretic transfer of proteins to nitrocellulose sheets and subsequent detection by immunological procedures. The proteins identified include a number of serum proteins as well as prostatic acid phosphatase and creatine kinase. Proteolytic events shown to occur during the liquefaction of semen that occurs early after collection indicate the importance of carefully controlled collection and preparation methods for clinical evaluation of seminal plasma. Ethylenediaminetetraacetic acid and phenylmethylsulfonyl fluoride inhibit this proteolysis.

Concanavalin A↗
PubMed1981

The TYCHO system for computer analysis of two-dimensional gel electrophoresis patterns.

We describe here a computer system for the analysis of high-resolution two-dimensional gel-electrophoresis patterns, with some initial applications. The system (called TYCHO) comprises programs for image acquisition, background subtraction and smoothing, spot detection, gaussian spot modeling, and pattern matching and comparison. It is based on a conventional minicomputer, but makes extensive use of a high-speed array processor in the image-processing and -modeling steps. Used in concert with the ISO-DALT two-dimensional electrophoresis system (Anal. Biochem. 85:331-354, 1978), TYCHO allows quantitative measurement of hundreds of proteins in complex biological samples, and constitutes the initial data-reduction system required for work towards a Human Protein Index.

Blood Proteins↗
PubMed1981

Muscle protein analysis. III. Analysis of solubilized frozen-tissue sections by two-dimensional electrophoresis.

Proteins from frozen histological sections of human muscle were analyzed by two-dimensional gel electrophoresis. Patterns so obtained were identical to those from whole homogenates of muscle prepared from frozen tissue powders that had much higher protein concentrations. To increase the number of proteins visible on gels of samples low in protein content, the gels were silver stained, or the proteins were labeled with [14C]iodoacetamide before electrophoresis and the gels were fluorographed. The latter method allow use of a single frozen-tissue section for two-dimensional electrophoretic analysis and brings the technique closer to practicable clinical use.

Actins↗
PubMed1980

Synthesis and biological activity of 5-(4-chlorobenzoyl)-4-(hydroxymethyl)-1-methyl-1H-pyrrole-2-acetic acid, a major metabolite of zomepirac sodium.

5-(4-Chlorobenzoyl)-4-(hydroxymethyl)-1-methyl-1H-pyrrole-2-acetic acid (2), the major oxidative metabolite of zomepirac (1), was synthesized starting with ethyl 5-(4-chlorobenzoyl)-1,4-dimethyl-1H-pyrrole-2-acetate (3), the ethyl ester of 1. Compound 3 was oxidized with selenium dioxide to afford the alpha-oxoester, 5. Bromination of 5 with N-bromosuccinimide produced bromomethylpyrrole 7, and reaction of 7 with acetate produced by corresponding acetoxymethylpyrrole 8. Hydrogen sulfide effected the selective reduction of the side-chain carbonyl group if 8 to give 9. Saponification of 9 gave the title compound, 2. Synthetic 2 was identical with the isolated metabolite of zomepirac (1). Biological testing revealed that the metabolite was essentially devoid of the biological activity associated with zomepirac.

Analgesics↗
PubMed1980

C19 steroidal precursors of estrogens.

The pathways of biosynthesis of the estrogens from C19 steroids has been examined by in vitro kinetic experiments using human placental microsomes and two precursors, each labeled with a different radioisotope. The results indicate that under the conditions employed, androstenedione is an obligatory intermediate in the conversion of 3 beta-hydroxy-5-androsten-17-one (dehydroisoandrosterone) or its sulfate, dehydroisoandrosterone sulfate, into estrone and estradiol. The same result was obtained when the microsomal fraction was replaced by either a 1000 x g supernatant preparation from human placenta or a homogenate of placental mitochondria. If alternative pathways involving C19-hydroxylated derivatives of dehydroisoandrosterone or its sulfate exist, they appear to be of minor quantitative significance.

Androstenedione↗
PubMed1980

Muscle protein analysis. II. Two-dimensional electrophoresis of normal and diseased human skeletal muscle.

We used high-resolution two-dimensional electrophoresis to analyze the major proteins of normal and pathological human-muscle samples. The normal human-muscle pattern contains four myosin light chains: three that co-migrate with the myosin light chains from rabbit fast muscle (extensor digitorum longus), and one that co-migrates with the light chain 2 from rabbit slow muscle (soleus). Of seven Duchenne muscular dystrophy samples, four yielded patterns with decreased amounts of actin and myosin relative to normal muscle, while three samples gave patterns comparable to that for normal muscle. Six samples from patients with myotonic dystrophy also gave normal patterns. In nemaline rod myopathy, in contrast, the pattern was deficient in two of the fast-type myosin light chains.

Adult↗