Acid phosphatase in eggs of the zebrafish,Brachydanio rerio

Summary 2 isozymes of acid phosphatase have been identified by polyacrylamide disc gel electrophoresis in the ovary and mature, unfertilized eggs ofB. rerio. Histochemically, the enzyme appears to be localized in preyolk bodies of previtellogenic oocytes and in yolk platelets of vitellogenic and postvitellogenic oocytes. The contents of the cortical granules at all stages of oocyte differentiation were acid phosphatase negative.     

Alkaline phosphatase isozymes in cultured human cancer cells

Alkaline phosphatase, an ubiquitous enzyme is known to exist in several isozymic forms. At least three different isozymes have now been identified in humans. Alkaline phosphatase isozymes are among the substances synthesized ectopically by a variety of human tumors and many continuous cell lines derived from different cancers have retained the capacity to produce these membrane-located glycoproteins. This paper reviews the identification of alkaline phosphatase isozymes in cultured tumor cells and relates these findings with recent developments concerning these cell membrane located glycoproteins.     

Alkaline phosphatase isozymes in cultured human cancer cells

Summary Alkaline phosphatase, an ubiquitous enzyme is known to exist in several isozymic forms. At least three different isozymes have now been identified in humans. Alkaline phosphatase isozymes are among the substances synthesized ectopically by a variety of human tumors and many continuous cell lines derived from different cancers have retained the capacity to produce these membrane-located glycoproteins. This paper reviews the identification of alkaline phosphatase isozymes in cultured tumor cells and relates these finding with recent developments concerning these cell membrane located glycoproteins.     

Serological and genetical studies on the evolution of substrate specificity of flavone glycosyltransferase genes inSilene

Summary The variation in flavone glycosylation patterns inSilene is the result of the expression of six genetic loci, which control either the presence of allozymes differing in substrate specificity or isozymes regulated differently during development. Serological studies showed that at least three of these six loci are evolutionarily related. The genetic mechanisms leading to these complicated variation patterns and the role of this polymorphism for the plant in its interaction with the environment are discussed.     

Differential expression of isozymes in relation to organogenesis in two closely related species

Summary Two closely related species of Gentianaceae were established in tissue culture and transferred to a regeneration medium containing 2,4-D. Even though the age of cultures and all the environmental conditions were identical, only one of them expressed a clear morphogenic potential, and the differences are expressive in the isozyme banding patterns observed for creatine phosphokinase, lactate dehydrogenase and indoleacetaldehyde dehydrogenase. No differences in alkaline phosphatase isozymes were detected.This work was supported in part by the National Science Foundation (No. DCB 8 611 889) USA with a grant to the senior author.     

Hepatic acid hydrolases of albino rats,Mastomys natalensis and albino mice duringPlasmodium berghei infection

Summary Changes in liver acid hydrolase activities during the infection of albino rats,Mastomys or mice withPlasmodium berghei are described. B-Glucosidase, B-galactosidase and N-acetyl-B-D-glucosaminidase exhibited widely different responses with acid phosphatase and cathepsin-B the least responsive and are likely to be causally related to immunity of animals.     

A testis-specific lactate dehydrogenase in the pipid frog,Hymenochirus boettgeri

Summary Lactate dehydrogenase zymograms of mature testes ofHymenochirus boettgeri show in addition to the five isozymes composed of LDH-A and LDH-B subunits, a second 5-band system which is due to isozymes formed between LDH-A and a third subunit, LDH-C. These testis-specific LDH-C isozymes appear around 6 months after metamorphosis indicating that their expression is correlated with sexual maturity as is generally the case in mammals. This is the first report of a testis-specific LDH isozyme in a lower tetrapod; such isozymes have hitherto only been reported in mammals and in the pigeon.This study was supported by a grant from the Fond National Suisse de la Recherche Scientifique to H.R.K. We thank M. Sateur and F. Bourquin for their technical help.     

Electrophoretic variation in esterases in 3 wild-type and respective mutant strains ofAspergillus flavus

Summary Wild-type strains and auxotrophic mutants ofAspergillus flavus, differing regarding aflatoxin production, were tested for esterases isozymes. Esterases variation was found in all strains used, and a possible correlation between the pattern of esterase bands and aflatoxin production is suggested.Acknowledgment. The authors are thankful to the National Council for the development of Science and Technology (CNPq) and Assistance to Research for São Paulo State (FAPESP) for financial assistance.     

Genetic control of LDH isozymes in theRana esculenta complex

Summary Studies of LDH isozymes in the European green frogs showed that the synthesis of the B subunits is controlled by 3 alleles at a single genetic locus. The genetic evidence supports the hypothesis thatRana esculenta is the hybrid ofR. lessonae x R. ridibunda.This work was supported by grants from the Schweizerischer Nationalfonds, the Georges und Antoine Claraz-Schenkung and the Karl Hescheler-Stiftung.     

Protein kinase C regulation of GABAA receptors

Pharmacological studies with drugs that activate or inhibit several protein kinase C (PKC) isozymes have identified the PKC family of serine-threonine kinases as important in the regulation of -aminobutyric acid type A (GABA_A) receptor function. PKC modulates GABA_A receptor surface density, chloride conductance and receptor sensitivity to positive allosteric modulators such as neurosteroids, ethanol, benzodiazepines and barbiturates. Recent studies using PKC isozyme-selective reagents and gene-targeted mice have begun to identify critical roles for three isozymes, PKCII, PKC and PKC, in various aspects of GABA_A receptor regulation. Progress in this field touches upon therapeutic areas that are of great clinical importance such as anxiety and addiction. Increased understanding of how PKC regulates GABA_A receptors and which PKC isozymes are involved holds promise for development of new treatments for diverse neuropsychiatric disorders.Received 2 August 2004; received after revision 17 August 2004; accepted 21 August 2004     

Histochemical localization of alkaline and acid phosphatase activities in the skin ofMystus (Mystus) vittatus Bl. (Siluriformes)

Summary An attempt has been made to localize alkaline and acid phosphatase activities in the skin ofMystus vittatus by using histochemical techniques. The alkaline phosphatase activity is found in metabolically active cells such as basal columnar cells, mucous cells and polygonal support cells. The acid phosphatase activity is intense in the outermost squamous support cells and in the basal columnar cells. These activities have been correlated with some physiological functions of the epidermis.Acknowledgment. We are thankful to P. Vishwanatham, Government College, Mhow, and Dr R.S. Shrivastava, Holkar Science College, Indore, for providing laboratory facilities and to the Council of Scientific and Industrial Research, New Delhi, for a fellowship for M.S.     

Genetic analysis of the kinome and phosphatome in cancer

Protein phosphorylation is a well-characterized biochemical process for reversible regulation of protein activity. Protein kinases and protein phosphatases are the key complementary players in this process, and through their coordinated activity cell homeostasis is tightly controlled. If these enzymes display aberrant activity, cells may undergo unrestrained growth, thus giving rise to complex diseases such as cancer. The technological platform gathered during the Human Genome Project recently allowed the systematic identifi cation of the genetic alterations present in the kinase (the kinome) and the phosphatase (the phosphatome) gene families. These studies suggest that most if not all human tumors carry genetic alterations in at least one phosphatase or kinase gene. Here we integrate the biochemical knowledge on the properties of these molecules with the information collected through their systematic genetic analysis in cancer. We also analyze why the molecular profi ling of the kinome and phosphatome in individual cancers is revolutionizing basic and clinical oncology.Received 13 May 2005; received after revision 30 May 2005; accepted 22 June 2005     

Phosphomonoesterases in the 2 sexes of the root-knot nematode,Meloidogyne lucknowica Singh, 1969

Summary The 2 phosphomonoesterases of the root-knot nematode were colorimetrically determined. Alkaline phosphatase activity was observed to be lower than the acid phosphatase activity. Sex related trends were clearly seen in the enzyme levels of the 2 sexes of thenematode. Alkaline phosphatase level differed 28.76%, while acid phosphatase level differed 60.36% in the 2 sexes.The authors are grateful to the Council of Science and Technology U.P. for the grant of a Research Scheme for these studies.     

Dissociation-constants of metal-ion-complexes with alkaline phosphatase from pig kidney

Summary Using metal-ion buffers it was possible to remove Zn²⁺, Mg²⁺ and Mn²⁺ ions of pig kidney alkaline phosphatase reversibly. The dissociation constants obtained are K_EMg:4·10^–7 M, K_EMn:4·10^–8 M and K_EZn:8·10^–13 M (22°C, pH:9.6, :0.07).Acknowledgement: The authors thank Dr.H. U. Wolf for helpful suggestions and valuable discussion and MissH. Köth for technical assistance.     

Lactate dehydrogenase isozymes of mouse epidermis

Summary Five isozymes of LDH are demonstrable in the epidermis of the ear pinnae, hind feet, trunk dorsa, and tails of adult C57BL, C57HR, and C3HB mice by polyacrylamide gel electrophoresis. LDH-5 activity predominates in electropherograms. The ratio of LDH-1 to LDH-5 is greater in the epidermis of ear pinna and trunk dorsum than in that of tail and hind foot. The region-specific patterns of epidermal LDH isozymes are not correlated with melanin pigmentation or ‘hairiness’ of the skin. Supported by PHS Research Grant No. CA-06097 from the National Cancer Institute, United States Public Health Service.     

Increase in acid phosphatase activity during fertilization of a teleost egg

Summary Acid phosphatase activity was localized at the periphery of the yolk globules in the egg of the teleost,Catostomus commersoni. In addition, a 3-fold increase in acid phosphatase activity was associated with fertilization and pronucleus formation.I thank Dr C. W. Huver for use of facilities.     

Expression of the Stp1 LMW-PTP and inhibition of protein CK2 display a cooperative effect on immunophilin Fpr3 tyrosine phosphorylation and <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> growth

Although the yeast genome does not encode bona fide protein tyrosine kinases, tyrosine-phosphorylated proteins are numerous, suggesting that besides dual-specificity kinases, some Ser/Thr kinases are also committed to tyrosine phosphorylation in Saccharomyces cerevisiae. Here we show that blockage of the highly pleiotropic Ser/Thr kinase CK2 with a specific inhibitor synergizes with the overexpression of Stp1 low-molecular-weight protein tyrosine phosphatase (PTP) in inducing a severe growth-defective phenotype, consistent with a prominent role for CK2 in tyrosine phosphorylation in yeast. We also present in vivo evidence that immunophilin Fpr3, the only tyrosine-phosphorylated CK2 substrate recognized so far, interacts with and is dephosphorylated by Spt1. These data disclose a functional correlation between CK2 and LMW-PTPs, and suggest that reversible phosphorylation of Fpr3 plays a role in the regulation of growth rate and budding in S. cerevisiae.Received 15 January 2004; received after revision 20 February 2004; accepted 4 March 2004     

Optimization of the cellular import of functionally active SH2-domain-interacting phosphopeptides

Phosphopeptides interacting with src homology 2 (SH2) domains can activate essential signaling enzymes in vitro. When delivered to cells, they may disrupt protein-protein interactions, thereby influencing intracellular signaling. We showed earlier that phosphopeptides corresponding to the inhibitory motif of Fcγ receptor IIb and a motif of the Grb2-associated binder 1 adaptor protein activate SH2-containing tyrosine phosphatase 2 in vitro. To study the ex vivo effects of these peptides, we have now compared different methods for peptide delivery: (i) permeabilization of the target cells and (ii) the use of cell-permeable vectors, which are potentially able to transport biologically active compounds into B cells. We found octanoyl-Arg₈ to be an optimal carrier for the delivery of phosphopeptides to the cells. With this strategy, the function of cell-permeable SHP-2-binding phosphopeptides was analyzed. These peptides modulated the protein phosphorylation in B cells in a dose- and time-dependent manner. Received 27 July 2006; received after revision 4 September 2006; accepted 18 September 2006     

Diagnostic alleles and systematics in termite species of the genusReticulitermes in europe

Summary It was possible to separate species and complexes of species in theReticulitermes genus of Western Europe using esterase 3 and acid phosphatase 2.     

Isozymes of cathepsin B1 in developing human placenta

Summary Cathepsin B1 was purified from human placentas of different gestational ages and analyzed by isoelectric focusing on polyacrylamide gel. The enzyme was shown to consist of 3 or 4 isozymes with pI values between 5.1 and 5.7.