SVIBOR - Project code: 1-07-196

MINISTRY OF SCIENCE AND TECHNOLOGY

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tel.: +385 1 459 44 44, fax: +385 1 459 44 69
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Project code: 1-07-196


PROTEOLYTIC ENZIMES: PROPERTIES, STRUCTURE AND FUNCTION


Main researcher: VITALE, LJUBINKA (52520)


Assistants
Type of research: basic
Duration from: 01/01/91. to 12/31/95.
Papers on project (total): 23
Papers on project quoted in Current Contents: 6
Institution name: Institut "Ruđer Bošković", Zagreb (98)
Department/Institute: Department of Organic Chemistry and Biochemistry
Address: Bijenička c. 54,
City: 10000 - Zagreb, Croatia
Communication
Phone: 385 (41) 424-987
Fax: 385 (41) 425-647
Summary: (1) To broaden the knowledge of proteolytic enzymes systems in human blood cells and to elucidate the role of aminopeptidases, the properties, localization and structure of these enzymes will be studied. In particular, aminopeptidases which prefer arginine and those which prefer methionine at the NH2-end of the substrate will be investigated. The enzymes will be purified and characterized as catalysts and protein molecules as well. Further, their distribution in different tissues and cells and within the cell itself will be followed. Thus, aminopeptidases localization with respect to different types of polymorphonuclear leukocyte granules will be determined and concomitantly the granules better defined. Enzymes from different sources will be compared. (2) Proteolytic and other hydrolytic enzymes from industrial microorganism Streptomyces rimosus will also be studied. Procedures for their isolation will be designed, and their properties and partial structure determined. Immunochemical assay for determination of relatedness of the same type enzymes from different strains and species of streptomycetes will be developed, and its applicability for taxonomy of these bacteria evaluated. (3) As a part of the project a care for establishing a facility for determination of the primary structure of proteins and peptides was foreseen (instrument, running conditions, stuff training). The facility would be available to all those in Croatia who have a need for determination of amino acids sequences.

Keywords: proteolytic enzymes, aminopeptidases, erythrocytes, polymorphonuclear leukocytes, granules, Streptomyces rimosus, amylase, nuclease, enzyme isolation, enzyme localization, immunochemical assays

Research goals: One of the project aims is to determine a number and type of enzymes responsible for the aminopeptidase activities found in human blood cells, to determine their properties and specificity and to establish to which subcellular compartment a particular aminopeptidase is confined. Further, to determine relatedness of aminopeptidases from different cells and their distribution among other tissues. For better understanding of structure - mechanism of action relation, first a partial and later on a complete primary structure of one of the isolated enzymes will be determined, providing necessary facilities will be available. The research should result with new data about aminopeptidases and their physiological role, about blood cells and their differentiation, and in particular about the function of polymorphonuclear leukocyte granules. For microbial enzymes the aim is to find a suitable preparation procedure, to evaluate their application possibility and to elaborate an immunochemical method for determination of some of peptidases and other hydrolases, which could at the same time diferentiate strains and species of streptomycetes. For one of the enzymes isolated from S. rimosus partial primary structure on protein and on gene level would be determined, hoping to get an insight into its postsynthetic modifications. The new data on hydrolytic enzymes and on streptomycetes are expected, whereas the procedures for isolation and determination of enzymes might have a practical value. The aim of the project is also a development of methods for separation and analysis of proteins in general.

COOPERATION - INSTITUTIONS


  1. Name of institution: Odsek za biokemijo in molekularno biologijo Institut "Jožef Stefan"
    Type of institution: University/Faculty
    City: 61000 - Ljubljana, Slovenija

  2. Name of institution: Herbos d.d.
    Type of institution: State institute
    City: 44000 - Sisak, Croatia
Other information about the project.
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Last update: 02/22/94
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