Harry Harris

Bio: Harry Harris is an academic researcher from University College London. The author has contributed to research in topics: Isozyme & Alkaline phosphatase. The author has an hindex of 54, co-authored 143 publications receiving 11630 citations.

Isolation and characterization of a cDNA encoding a human liver/bone/kidney-type alkaline phosphatase.

Abstract: Alkaline phosphatases (ALPs) [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] isolated from human liver, bone, and kidney (L/B/K) exhibit very similar biochemical and immunologic properties that differentiate them from other human ALPs, such as those characteristically found in placenta and intestine. Despite their similarities, the L/B/K ALPs produced in different tissues show slight physical differences. To examine structural and evolutionary relationships between the various ALPs, a cDNA corresponding to L/B/K ALP mRNA has been isolated. A lambda 11 cDNA expression library was constructed using poly(A) RNA from the osteosarcoma cell line Saos-2 and screened with anti-liver ALP antiserum. The 2553-base-pair cDNA contains an open reading frame that encodes a 524 amino acid polypeptide with a predicted molecular mass of 57.2 kDa. This ALP precursor protein contains a presumed signal peptide of 17 amino acids followed by 37 amino acids that are identical to the amino-terminal sequence determined from purified liver ALP. In addition, amino acid sequences of several CNBr peptides obtained from liver ALP are found within the cDNA-encoded protein. The deduced L/B/K ALP precursor polypeptide shows 52% homology to human placental ALP and 25% homology to Escherichia coli ALP precursor polypeptides. Sixty percent nucleotide homology exists between the human L/B/K and placental cDNAs over the protein coding regions. The 5' and 3' untranslated regions of the L/B/K ALP cDNA, 176 and 805 base pairs, respectively, show no homology to the corresponding regions of placental ALP cDNA.

Esterase D: a new human polymorphism

Abstract: Tashian (1 961, 1969) demonstrated several different esterases in human red cells by electrophoresis. Azo dye coupling techniques were used to locate the esterase isozymes. Three main groups of esterases were defined on the basis of their electrophoretic properties, substrate specificities and inhibition characteristics, and were referred to as the A, B and C esterases. Isozymes of carbonic anhydrase, which also has esteratic activity, were also demonstrated using essentially similar techniques. Fig. 1, which is based on Tashian's work, shows a diagrammatic representation of the various red cell esterase isozymes after electrophoresis of haemolysates in borate starch gels at pH 8.0-8-6. The A esterases are subdivided into three sets of isozymes, A,, A, and As, which differ electrophoretically and in their storage properties and inhibition characteristics ; also the As isozymes have a larger molecular size than A, or A,. Inherited variants of these isozymes are rare in man but two different variant phenotypes have been reported (Tashian & Shaw, 1962; Tashian, 1965). They are of particular interest because each of the three subgroups of A esterase are affected in the variant isozyme patterns suggesting that A,, A, and As contain a common subunit. Family studies on one of the variants indicated that this esterase A locus is autosomal. Esterase B is primarily a butyryl esterase and no variants have been reported in population surveys to date. Esterase C is a rather weakly staining invariate acetyl esterase. Several different variants of the carbonic anhydrase isozyme CA I (or B) have been recorded in the literature (see Tashian, 1969) but they are all individually rare except in certain cases where they have been identified in small isolated populations, e.g. the CA I, variant in Guam and Saipan. The CA 1 locus is autosomal and the variants do not affect CA I1 (or C). The latter is determined at a separate autosomal locus and recent studies (Moore, Funakoshi & Deutsch, 1971) using an immunological detection technique have led to the recognition of a genetic polymorphism of CA 11 in Blacks. Another esterase which occurs in red cells is acetylcholinesterase. This is membrane bound and not therefore readily detected by electrophoresis of simple lysates. The present paper describes yet a further human red cell esterase. It will be referred to as esterase D (Es D). It was discovered by the use of the fluorogenic substrates, 4-methyl-umbelliferyl acetate and 4-methyl-umbelliferyl butyrate, for the detection of esterase isozymes after starch gel electrophoresis. The isozymes of this new esterase are different from the red cell A, B aiid C esterases, the carbonic anhydrase isozymes, and from acetylcholinesterase in their

Developmental changes and polymorphism in human alcohol dehydrogenase

Abstract: Ann. H u m . Genet., Lond. (1971), 34, 251 Printed in Great Britain Developmental changes and polymorphism in human alcohol dehydrogenase BY MOYRA SMITH, D. A. HOPKINSON AND HARRY HARRIS M.R.C. Human Biochemical Genetics Unit, Galton Laboratory, University College London I n man, alcohol dehydrogenase (alcohol: NAD oxidoreductase E.C. 1 . 1 . 1 . 1 ) occurs princi- pally in liver, though low levels of activity have aIso been found in lung, kidney and the gastro- intestinal tract (Moser, Papenberg & von Wartburg, 1968). Evidence for at least three distinct isozymes has been obtained by chromatography of liver extracts on CM cellulose (Blair & Vallee, 1966) and also by electrophoresis (Moser et al. 1968; Pikkarainen & Raiha, 1969; Murray & Motulsky, 1970). Von Wartburg, Papenberg & Aebi (1965) reported that certain individuals have an atypical form of alcohol dehydrogenase associated with an increased level of activity. The usual and atypical forms of the enzyme were shown to differ markedly in pH activity curves with ethanol as substrate. The pH optimum for the usual form was found to be pH 10.8 and for the atypical form pH 8.5. The enzymes also differed in the relative rates at which they oxidized various other alcohols, and in the degree of inhibition produced by various metal binding agents. On the other hand no significant differences were observed in Michaelis constants for the substrates ethanol or acetaldehyde or for the corresponding coenzymes NAD or NADH. Also the pH activity curve with acetaldehyde as substrate was essentially the same for both enzymes, having an optimum at pH 6.0-6.5. A simple screening test to distinguish the usual from the atypical enzyme in crude liver homo- genates was designed (von Wartburg et al. 1965). This involves determining the ratio of the activity at pH 11.0 to that at pH 8-8 with ethanol as substrate under standard conditions. The usual enzyme gives a value for this ratio greater than 1.0, and the atypical enzyme less than 1.0. In a survey of 59 liver samples from different individuals in Switzerland, 12 were found to have the atypical alcohol dehydrogenase, and in another series of 50 individuals from London, 2 were found to be atypical (von Wartburg & Schiirch, 1968). The atypical enzyme occurred in indi- viduals varying from 16 to 82 years of age. Pikkarainen & Raiha (1967) reported that alcohol dehydrogenase activity in liver is low during foetal life and reaches adult levels about 5 years after birth. Changes in electrophoretic pattern have also been noted during development (Pikkarainen & Raiha, 1969; Murray & Motulsky, 1970). In the earliest stages only a single isozyme is observed but later further iso- zymes appear. I n adult liver individual variations in the relative contribution of the different isozymes to the total activity have been noted (von Wartburg & Schiirch, 1968), but no clear electrophoretic differences between the usual and atypical alcohol dehydrogenases as determined by the ratio of activity at pH 11.0 and pH 8.8 were detected. The present paper is concerned with a study of human alcohol dehydrogenase in which liver, lung, kidney and intestinal material from foetuses, infants and adults has been examined. The en- zyme has been investigated both by spectrophotometric assay at different pH’s and by starch-gel

Advances in Human Genetics

Abstract: This book has five chapters covering peroxisomal diseases, X-linked immunodeficiencies, genetic mutations affecting human lipoproteins and their receptors and enzymes, genetic aspects of cancer, and Gaucher disease. The chapter on peroxisomes covers their discovery, structure, functions, disorders, etc. The chapter on X-linked immunodeficiencies discusses such diseases as agammaglobulinemia, severe combined immunodeficiency, Wiskott-Aldrich syndrome, animal models, linkage analysis, etc. Apolipoprotein formation, synthesis, gene regulation, proteins, etc. are the main focus of chapter 3. The chapter on cancer covers such topics as oncogene mapping and the molecular characterization of some recessive oncogenes. Gaucher disease is covered from its diagnosis, classification, and prevention, to its organ system involvement and molecular biology.

Analysis of molecular variance inferred from metric distances among DNA haplotypes: application to human mitochondrial DNA restriction data.

Abstract: We present here a framework for the study of molecular variation within a single species. Information on DNA haplotype divergence is incorporated into an analysis of variance format, derived from a matrix of squared-distances among all pairs of haplotypes. This analysis of molecular variance (AMOVA) produces estimates of variance components and F-statistic analogs, designated here as phi-statistics, reflecting the correlation of haplotypic diversity at different levels of hierarchical subdivision. The method is flexible enough to accommodate several alternative input matrices, corresponding to different types of molecular data, as well as different types of evolutionary assumptions, without modifying the basic structure of the analysis. The significance of the variance components and phi-statistics is tested using a permutational approach, eliminating the normality assumption that is conventional for analysis of variance but inappropriate for molecular data. Application of AMOVA to human mitochondrial DNA haplotype data shows that population subdivisions are better resolved when some measure of molecular differences among haplotypes is introduced into the analysis. At the intraspecific level, however, the additional information provided by knowing the exact phylogenetic relations among haplotypes or by a nonlinear translation of restriction-site change into nucleotide diversity does not significantly modify the inferred population genetic structure. Monte Carlo studies show that site sampling does not fundamentally affect the significance of the molecular variance components. The AMOVA treatment is easily extended in several different directions and it constitutes a coherent and flexible framework for the statistical analysis of molecular data.

Construction of a genetic linkage map in man using restriction fragment length polymorphisms.

Abstract: We describe a new basis for the construction of a genetic linkage map of the human genome. The basic principle of the mapping scheme is to develop, by recombinant DNA techniques, random single-copy DNA probes capable of detecting DNA sequence polymorphisms, when hybridized to restriction digests of an individual's DNA. Each of these probes will define a locus. Loci can be expanded or contracted to include more or less polymorphism by further application of recombinant DNA technology. Suitably polymorphic loci can be tested for linkage relationships in human pedigrees by established methods; and loci can be arranged into linkage groups to form a true genetic map of "DNA marker loci." Pedigrees in which inherited traits are known to be segregating can then be analyzed, making possible the mapping of the gene(s) responsible for the trait with respect to the DNA marker loci, without requiring direct access to a specified gene's DNA. For inherited diseases mapped in this way, linked DNA marker loci can be used predictively for genetic counseling.

Hypervariable 'minisatellite' regions in human DNA.

Abstract: The human genome contains many dispersed tandem-repetitive 'minisatellite' regions detected via a shared 10-15-base pair 'core' sequence similar to the generalized recombination signal (chi) of Escherichia coli. Many minisatellites are highly polymorphic due to allelic variation in repeat copy number in the minisatellite. A probe based on a tandem-repeat of the core sequence can detect many highly variable loci simultaneously and can provide an individual-specific DNA 'fingerprint' of general use in human genetic analysis.

The many faces of publish/subscribe

Abstract: Well adapted to the loosely coupled nature of distributed interaction in large-scale applications, the publish/subscribe communication paradigm has recently received increasing attention. With systems based on the publish/subscribe interaction scheme, subscribers register their interest in an event, or a pattern of events, and are subsequently asynchronously notified of events generated by publishers. Many variants of the paradigm have recently been proposed, each variant being specifically adapted to some given application or network model. This paper factors out the common denominator underlying these variants: full decoupling of the communicating entities in time, space, and synchronization. We use these three decoupling dimensions to better identify commonalities and divergences with traditional interaction paradigms. The many variations on the theme of publish/subscribe are classified and synthesized. In particular, their respective benefits and shortcomings are discussed both in terms of interfaces and implementations.