Abstract: The present invention provides a human prostate-associated protease (HUPAP) and polynucleotides which identify and encode HUPAP. The invention also provides expression vectors, host cells, antibodies, antagonists, and antisense molecules. The invention also provides methods for treating disorders associated with expression of HUPAP.

Abstract: The present invention relates generally to proteinase inhibitors, a precursor thereof and to genetic sequences encoding same. More particularly, the present invention relates to a nucleic acid molecule comprising a sequence of nucleotides which encodes or is complementary to a sequence which encodes a type II serine proteinase inhibitor (PI) precursor from a plant wherein said precursor comprises at least three PI monomers and wherein at least one of said monomers has a chymotrypsin specific site and at least one other of said monomers has a trypsin specific site.

Abstract: The invention relates to a method for the renaturation of denatured proteins in which they are treated with a renaturant which has on vicinal carbon atoms a hydroxyl group and at least one fluorine atom.

Abstract: Novel herpes viral protease crystalline structures are identified which have an active site formed by the three amino acids Ser, His and His. Also disclosed are methods of identifying inhibitors of these proteases and/or active sites.

Abstract: A method is described for a simple, fast and efficient synthesis of homopolymers and copolymers by the enzymatic ring opening polymerization of lactones and lactides. The enzyme used is an ion paired protease. The advantage of this enzymatic system is in using small amount of enzyme per monomer and lower reaction time. Homopolymers and copolymers are synthesized with molecular weights between 1000 and 4600 daltons, and dispersity as low as 1.1. The monomer conversion after 4 days, for reactions catalyzed by protease S, has reached 100%. Different initiators are used to control the rate and degree of polymerization. Synthesis of block copolymers with defined block size and crystallinity are described in this invention. These biodegradable and bioerodable polyesters and copolyesters with controlled molecular weight, dispersity and crystallinity have applications in medical, drug, cosmetic and food industries.

Abstract: Trypsin (trypsinogen) may be produced in a filamentous fungus by transforming a filamentous fungus with a vector comprising a DNA sequence encoding protrypsin or a derivative thereof N-terminally fused to a DNA sequence encoding a signal peptide, culturing the transformed filamentous fungus in a suitable culture medium to produce trypsinogen and recovering trypsinogen and/or trypsin from the medium.

Abstract: Serine protease inhibitors are provided comprising compounds having a P site binding moiety and a divalent cation(s) chelating moiety which are pre-prepared as divalent cation(s) complexes or combined with the serine protease in the presence of divalent cation(s). The compounds are shown to have high inhibitory activity when complexed to divalent cation(s) and find use in various processes associated with serine protease isolation and inhibition.

Abstract: Serine protease inhibitors are provided comprising compounds having a P site binding moiety and a divalent cation(s) chelating moiety which are pre-prepared as divalent cation(s) complexes or combined with the serine protease in the presence of divalent cation(s). The compounds are shown to have high inhibitory activity when complexed to divalent cation(s) and find use in various processes associated with serine protease isolation and inhibition.

Abstract: Analogs of the serine protease inhibitors in which the amino acid sequence is varied slightly are disclosed, which analogs variously show improved properties including improved resistance to oxidative inactivation, improved ability to inhibit pancreatic elastase, improved ability to inhibit cathepsin G, and improved ability to inhibit trypsin.

Abstract: A method of treating or preventing post-operative ileus in a mammalian subject is disclosed. The method involves administering to the subject, a pharmaceutically effective amount of a compound that is effective in (i) preventing mast cell degranulation, (ii) inhibiting tryptase and chymase, and (iii) antagonizing PAR-2. The treatment is based on the discoveries that proteinase-activated receptor 2 is expressed in colonic muscle cells, and that activation of PAR-2 inhibits colonic motility. The PAR-2 receptor is activated, at least in part, by tryptase and chymase, produced by infiltration and degranulation of mast cells.

Abstract: PEPZYMES.TM., chemically synthesized cyclic peptides, modeled on the active sites of naturally-occurring enzymes represented by chymotrypsin, trypsin, lysozyme, ribonuclease, urokinase, tissue plasminogen activator and their analogs are disclosed. The conformational constraints imposed on the peptide residues cause the amino acids of the peptide to assume a three-dimensional spatial relationship relative to the substrate that is essentially equivalent to that of the corresponding active site amino acids of the natural enzyme in its catalytically active state. The new cyclic peptides catalyze the same reaction as the native enzyme being modeled, but have amino acid sequences that are substantially shorter than the naturally-occurring enzymes, and do not occur in the same linear relationship in the naturally-occurring enzymes. Methods of producing PEPZYMES.TM. are described.

Abstract: The invention is directed to a pharmaceutical preparation with a modulatory effect on malignant tumors, which contains a combination of protease proenzymes and amylases in a ratio between 1:100 and 100:1 in enzymatic activity units, and a protease inhibitor. The invention is also directed to use of such a preparation to modulate the effects of malignant tumors in humans and animals.

Abstract: A method for making a collagen strengthened cellulosic sheet by providing a cellulosic pulp slurry; adding solubilized collagen to the pulp slurry, and mixing for a time effective for interaction of the cellulosic pulp slurry and solubilized collagen; forming the interacted cellulosic pulp slurry and solubilized collagen into a sheet; and drying the sheet; also, a method for using solubilized collagen for strengthening paper by mixing the solubilized collagen with a cellulosic pulp slurry; and making a cellulosic pulp product from the mixture and drying.

Abstract: This invention provides a nucleic acid sequence encoding a trypsin-like enzyme which can be present at the trachea of human lungs, and can selectively digest a synthetic substrate for trypsin and a synthetic substrate for thrombin, and fibrinogen; and a process for producing the trypsin-like enzyme by genetic engineering utilizing the nucleic acid sequence.

Abstract: A method for making a collagen strengthened cellulosic sheet by providing a cellulosic pulp slurry; adding solubilized collagen to the pulp slurry, and mixing for a time effective for interaction of the cellulosic pulp slurry and solubilized collagen; forming the interacted cellulosic pulp slurry and solubilized collagen into a sheet; and drying the sheet; also, a method for using solubilized collagen for strengthening paper by mixing the solubilized collagen with a cellulosic pulp slurry; and making a cellulosic pulp product from the mixture and drying.

Abstract: A method for making a collagen strengthened cellulosic sheet by providing a cellulosic pulp slurry; adding solubilized collagen to the pulp slurry, and mixing for a time effective for interaction of the cellulosic pulp slurry and solubilized collagen; forming the interacted cellulosic pulp slurry and solubilized collagen into a sheet; and drying the sheet; also, a method for using solubilized collagen for strengthening paper by mixing the solubilized collagen with a cellulosic pulp slurry; and making a cellulosic pulp product from the mixture and drying.

Abstract: The invention relates to serine protease variants derived from precursor serine proteases via recombinant and/or chemical methods to form protease variants having improved peptide ligase activity. The invention also includes novel ligation substrates which in combination with the serine protease variants and a second ligation substrate are capable of forming a ligation product. The invention also relates to methods for forming such ligation products and the products formed thereby.

Abstract: Organic enzyme solutions comprise an ion pair complex of an enzyme and a surfactant, and an organic solvent in which the enzyme-surfactant ion pair is dissolved. The solution has catalytic activity at least an order of magnitude greater than a suspension of an equal amount of the enzyme and organic solvent without ion pair complexes. The enzyme is preferably a hydrolase with an acyl transferase activity and the surfactant is Aerosol OT (sodium bis(2-ethylhexyl) sulfosuccinate) at low concentrations.

Abstract: A method for making a collagen strengthened cellulosic sheet by providing a cellulosic pulp slurry; adding solubilized collagen to the pulp slurry, and mixing for a time effective for interaction of the cellulosic pulp slurry and solubilized collagen; forming the interacted cellulosic pulp slurry and solubilized collagen into a sheet; and drying the sheet; also, a method for using solubilized collagen for strengthening paper by mixing the solubilized collagen with a cellulosic pulp slurry; and making a cellulosic pulp product from the mixture and drying.

Abstract: A method for making a collagen strengthened cellulosic sheet by providing a cellulosic pulp slurry; adding solubilized collagen to the pulp slurry, and mixing for a time effective for interaction of the cellulosic pulp slurry and solubilized collagen; forming the interacted cellulosic pulp slurry and solubilized collagen into a sheet; and drying the sheet; also, a method for using solubilized collagen for strengthening paper by mixing the solubilized collagen with a cellulosic pulp slurry; and making a cellulosic pulp product from the mixture and drying.

Abstract: The method of the invention provides for the formation of a recombinant polypeptide which has been modified at the C-terminal end through the use of a transpeptidation process. The method is suitable for modifying recombinant polypeptides of any source including those which may be commercially available, those derived from recombinant single copy or multicopy polypeptide constructs, or those derived from single or multicopy recombinant fusion protein constructs. The transpeptidation reaction involves contacting an endopeptidase enzyme with a recombinant polypeptide to substitute an addition unit, of one or more amino acids, for a leaving unit, linked to a core polypeptide through a cleavage site recognized by the endopeptidase enzyme. Recombinant polypeptides derived from multicopy polypeptide constructs may be cleaved from the multicopy polypeptide at the N-terminal and C-terminal ends and simultaneously under go substitution of the leaving unit by the desired addition unit.

Abstract: The present invention is related to a process for producing an active enzyme comprising fermenting the proform of the active enzyme in the present of a proteolytic enzyme different from the active enzyme and capable of converting the proenzyme into an active enzyme as well as to host cells, recombinant expression vectors and host cells suitable for use in the process.

Abstract: A method for making a collagen strengthened cellulosic sheet by providing a cellulosic pulp slurry; adding solubilized collagen to the pulp slurry, and mixing for a time effective for interaction of the cellulosic pulp slurry and solubilized collagen; forming the interacted cellulosic pulp slurry and solubilized collagen into a sheet; and drying the sheet; also, a method for using solubilized collagen for strengthening paper by mixing the solubilized collagen with a cellulosic pulp slurry; and making a cellulosic pulp product from the mixture and drying.

Abstract: A method for making a collagen strengthened cellulosic sheet by providing a cellulosic pulp slurry; adding solubilized collagen to the pulp slurry, and mixing for a time effective for interaction of the cellulosic pulp slurry and solubilized collagen; forming the interacted cellulosic pulp slurry and solubilized collagen into a sheet; and drying the sheet; also, a method for using solubilized collagen for strengthening paper by mixing the solubilized collagen with a cellulosic pulp slurry; and making a cellulosic pulp product from the mixture and drying.

Abstract: Serine protease inhibitors are provided comprising compounds having a P site binding moiety and a divalent cation(s) chelating moiety which are pre-prepared as divalent cation(s) complexes or combined with the serine protease in the presence of divalent cation(s). The compounds are shown to have high inhibitory activity when complexed to divalent cation(s) and find use in various processes associated with serine protease isolation and inhibition.

Abstract: The invention relates to a process for the preparation of N-protected-L-aspartyl-L-phenylalanine methyl ester by enzymatic coupling of an N-protected-L-aspartic acid and L- or DL-phenylalanine methyl ester in an aqueous solution with formation of a precipitate using a thermolysin-like protease enzyme, wherein a water-immiscible organic solvent is added to and blended with the reaction system during the formation of the precipitate in the course of the coupling reaction, which organic solvent has a relatively high affinity for the precipitate. The process is particularly suitable as a continuous process. The method results in lower enzyme deactivation during and after the enzymatic coupling reaction and in larger and thicker precipitated crystals.

Abstract: An active recombinant trypsin-like protease enzyme comprising the amino acid residues 25-224 of the amino acid sequence of SEQ ID NO:2, as well as a DNA construct encoding the enzyme and comprising the sequence of SEQ ID NO:1. The enzyme may be used as a detergent enzyme.

Abstract: The present invention relates to therapeutic methods wherein an actin-binding protein, preferably gelsolin or DBP, is administered to a patient with actin-containing clots in order to remove actin from the clots. The invention also relates to diagnostic methods in which actin is removed from an actin-containing clot in vitro and quantitated.

Abstract: The invention includes a method that provides a low cost aqueous solution of solubilized collagen by the steps of: (a) providing an aqueous ground slurry of insoluble collagen and adjusting the pH of said slurry to obtain activity for a proteolytic enzyme added in Step b; (b) adding said proteolytic enzyme to said pH adjusted slurry; (c) reacting the slurry and enzyme of Step b and/or recycled insoluble collagen and enzyme from Step e at a temperature, T, and for a time, t, effective for forming a solution increased in solubilized collagen; (d) adding additional water and insoluble collagen to said solution of Step c and mixing; (e) separating at least some of the solution of Step d containing solubilized collagen from insoluble collagen, whereby at least a portion of said insoluble collagen and proteolytic enzyme is recycled to Step c, and the separated solution containing solubilized collagen is withdrawn as product; an alternative embodiment provides for the direct production of solubilized collagen withou

Abstract: This invention particularly provides a novel polypeptide having high protease-inhibiting activity, preferably FXa-inhibiting activity, which comprises, at least as a part of the polypeptide, an amino acid sequence resulting from substitution of an amino acid for at least one amino acid in the following amino acid sequence (1), wherein the amino acid substitution is at least one substitution selected from the following substitution means (i) to (iii). It also provides a process for the production of the polypeptide, a novel DNA fragment encoding the polypeptide and a drug composition containing the same.Amino acid sequence (1) ##STR1## (i) Substitution of 15 position Gln counting from the N-terminus by an amino acid other than Gln.(ii) Substitution of 42 position Tyr counting from the N-terminus by an amino acid other than Tyr.(iii) Substitution of 7 position Arg counting from the N-terminus by an amino acid other than Arg.

Abstract: The invention includes an improved method for producing an aqueous solution of soluble collagen. The method comprises the steps of:(A) providing an aqueous ground slurry of insoluble collagen containing insoluble collagen at a first concentration and at a pH effective to obtain activity for a proteolytic enzyme added in step (B);(B) adding a proteolytic enzyme to said slurry;(C) reacting said slurry and enzyme at a temperature, T.sub.1, and for a time t.sub.1 effective for forming a slurry containing at least some soluble collagen;(D) diluting at least a portion of the slurry formed in step (C) with water to form a diluted slurry containing insoluble collagen at a second concentration; and(E) reacting said diluted slurry obtained in step (D) at a temperature, T.sub.2 and for a time t.sub.2, effective to produce a solution containing an increased amount of soluble collagen.

Abstract: A method for making a collagen strengthened cellulosic sheet by providing a cellulosic pulp slurry; adding solubilized collagen to the pulp slurry, and mixing for a time effective for interaction of the cellulosic pulp slurry and solubilized collagen; forming the interacted cellulosic pulp slurry and solubilized collagen into a sheet; and drying the sheet; also, a method for using solubilized collagen for strengthening paper by mixing the solubilized collagen with a cellulosic pulp slurry; and making a cellulosic pulp product from the mixture and drying.

Abstract: This invention particularly provides a novel polypeptide having high protease-inhibiting activity, preferably FXa-inhibiting activity, which comprises, an lease as a part of the polypeptide, an amino acid sequence resulting from substitution of an amino acid for at least one amino acid in the following amino acid sequence (1), wherein the amino acid substitution is at least one substitution selected from the following substitution means (i) to (iii). It also provides a process for the production of the polypeptide, a novel DNA fragment encoding the polypeptide and a drug composition containing the same.

Abstract: A process for preparing C-terminally amidated peptides, Peptide-NH.sub.2, is presented. In a first step, a substrate component is reacted with a nucleophile component in the presence of trypsin or a carboxypeptidase using as nucleophile a compound NH.sub.2 -R to form a first reaction product Peptide-NH-R. In a second step, the first reaction product is non-enzymatically chemically cleaved to form the C-terminally amidated product, Peptide-NH.sub.2. The substrate component is selected from a) peptide derivatives Peptide-X-Y, where X is an amino acid or peptide residue and Y is OH, OMe or C-terminal modification and c) C-terminally esterified peptides, Peptide-OR', where R' is alkyl, aryl, heteroaryl, or aralkyl. The nucleophile component is selected from ##STR1## wherein A-F and A'-E' are carbon atoms or up to two hetero atoms, Y is H, alkyl, aryl, aralkyl, oxo or carboxy, X.sup.1 -X.sup.5 are H or various substituents.

Abstract: Provide herein is a substantially pure gingipain-1 preparation, gingipain-1 being characterized as having an apparent molecular mass of 50 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and an apparent molecular mass of 44 kDa as estimated by gel filtration chromatography, said gingipain-1 having amidolytic and proteolytic activity for cleavage after arginine residues and having no amidolytic and/or proteolytic activity for cleavage after lysine residues, wherein the amidolytic and/or proteolytic activity is inhibited by cysteine protease group-specific inhibitors including iodoacetamide, iodoacetic acid, N-ethylmaleimide, leupeptin, antipain, trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane, TLCK, TPCK, p-aminobenzamidine, N-chlorosuccinamide, and chelating agents including EDTA and EGTA, wherein the amidolytic and/or proteolytic activity of said gingipain-1 is not sensitive to inhibition by human cystatin C, .alpha.2-macroglobulin, .alpha.

Abstract: A method for preparing protease-polyethylene glycol adducts is presented wherein the immobilized reversible inhibitor, benzamidine, prevents reaction of activated polyethylene glycol with the active site of the protease. Improved activity against macromolecular substrates is obtained compared to when the benzamidine is in solution during the conjugation reaction.

Abstract: The method of the invention provides for the formation of a recombinant polypeptide which has been modified at the C-terminal end through the use of a transpeptidation process. The method is suitable for modifying recombinant polypeptides of any source including those which may be commercially available, those derived from recombinant single copy or multicopy polypeptide constructs, or those derived from single or multicopy recombinant fusion protein constructs. The transpeptidation reaction involves contacting an endopeptidase enzyme with a recombinant polypeptide to substitute an addition unit, of one or more amino acids, for a leaving unit, linked to a core polypeptide through a cleavage site recognized by the endopeptidase enzyme. Recombinant polypeptides derived from multicopy polypeptide constructs may be cleaved from the multicopy polypeptide at the N-terminal and C-terminal ends and simultaneously under go substitution of the leaving unit by the desired addition unit.

Abstract: A composition for cleaning contact lenses includes a protease and a sugar in an amount effective to maintain substantially neutral pH in a borate buffer upon dissolution of the composition therein.

Abstract: A process for determining the concentration of cyclosporine in a sample, which comprises adding to the sample a predetermined amount of an isomerase which isomerizes N-terminal peptide to proline, then measuring the enzyme-catalyzed inhibition of the isomerization of a proline-containing substrate, and determining the concentration or cyclosporine from a calibration curve.

Abstract: The invention includes a method that produces a low cost aqueous solution of solubilized collagen by the steps of: (a) providing an aqueous ground slurry of insoluble collagen and adjusting the pH of said slurry to obtain activity for a proteolytic enzyme added in Step b; (b) adding said proteolytic enzyme to said pH adjusted slurry; (c) reacting the slurry and enzyme of Step b and/or recycled insoluble collagen and enzyme from Step e at a temperature, T, and for a time, t, effective for forming a solution increased in solubilized collagen; (d) adding additional water and insoluble collagen to said solution of Step c and mixing; (e) separating at least some of the solution of Step d containing solubilized collagen from insoluble collagen, whereby at least a portion of said insoluble collagen and proteolytic enzyme is recycled to Step c, and the separated solution containing solubilized collagen is withdrawn as product; an alternative embodiment provides for the direct production of solubilized collagen withou

Abstract: An isolated and purified substance called Acta having the following features: (a) a molecular weight of 60 kd to 70 kd in SDS-PAGE using 12.5% gel, (b) reacts with a monoclonal antibody which is secreted by hybridoma FERM BP-3482, (c) binds to chymotrypsin and (d) binds to DNA. Acta is used to diagnose cancer and Alzheimer's disease.

Abstract: This invention particularly provides a novel polypeptide having high protease-inhibiting activity, preferably FXa-inhibiting activity, which comprises, at least as a part of the polypeptide, an amino acid sequence resulting from substitution of an amino acid for at least one amino acid in the following amino acid sequence (1), wherein the amino acid substitution is at least one substitution selected from the following substitution means (i) to (iii). It also provides a process for the production of the polypeptide, a novel DNA fragment encoding the polypeptide and a drug composition containing the same.

Abstract: The present invention provides isolated DNA molecules comprising a DNA segment encoding a novel human amyloid protein precursor homologue and novel Kunitz-type inhibitors. Also provided are DNA constructs comprising a first DNA segment encoding a novel human amyloid protein precursor homologue or a novel Kunitz-type inhibitor wherein said first DNA segment is operably linked to additional DNA segments required for the expression for the first DNA segment, as well as host cells containing such DNA constructs and methods for producing proteins from the host cells.

Abstract: The present invention provides isolated DNA molecules comprising a DNA segment encoding a novel human amyloid protein precursor homolog and Kunitz-type inhibitor. Also provided are DNA constructs comprising a first DNA segment encoding a novel human amyloid protein precursor homolog or a Kunitz-type inhibitor wherein said first DNA segment is operably linked to additional DNA segments required for the expression for the first DNA segment, as well as host cells containing such DNA constructs and methods for producing proteins from the host cells.

Abstract: The invention relates to a process for preparing chirally uniform peptides. The aim of the invention is the preparation of chirally uniform peptides with the aid of proteolytic enzymes. The object comprises reacting alkyl esters of N-acylamino acids with proteases with amino acids, amino acid derivatives or peptides. The object is achieved by reacting an ester of an N-acylamino acid with a suitable amino acid derivative or peptide derivative with an unprotected N-terminal alphaamino group in frozen aqueous solution during catalysis with a serine protease or cysteine protease.

Abstract: Substituted isocoumarins, their use in inhibiting serine proteases with trypsin-like, chymotrypsin-like and elastase-like specificity and their roles as anti-inflammatory agents.

Abstract: The invention includes a method that produces a low cost aqueous solution of high molecular weight solubilized collagen by the steps of: (a) providing an aqueous ground slurry of insoluble collagen; (b) adjusting the water or solid content of the wet ground slurry whereby the insoluble collagen is at a concentration that promotes substantially maximum solubilized collagen concentration and molecular weight in a final product; (c) adjusting the pH of the slurry from Step b to obtain activity for a proteolytic enzyme added in Step d; (d) adding the proteolytic enzyme to the pH adjusted slurry and reacting at a temperature, T, and for a time, t, effective for forming high molecualr weight solubilized collagen from the insoluble collagen particles; (e) controlling the reaction conditions for obtaining a high concentration of soluble collagen and a high molecular weight of the solubilized collagen by simultaneously measuring the concentration of solubilized collagen and the molecular weight of the solubilized coll

Abstract: The design, and synthesis of peptide-based molecules termed helizymes which possess catalytic activity are described herein The catalytic molecules of the invention comprise a number of amphiphilic helical peptides, which interact via hydrophobic interactions and which are bonded at their carboxyl ends to a multifunctional base. Active site residues functional for catalysis are positioned within or at the N-termini of the helical peptides. The helizyme molecule adopts a conformation in aqueous medium such that a substrate binding pocket is formed and such that functional active site geometry results from the association of the helical peptides Specifically exemplified helizymes possess specificities and at least one catalytic activity of chymotrypsin, trypsin and acetylcholine esterase.

Abstract: The invention provides .alpha.-1-antichymotrypsin and protein preparations comprising human .alpha.-1-antichymotrypsin produced by E. coli cells transformed with a DNA sequence encoding human .alpha.-1-antichymotrypsin. The invention also provides methods for producing .alpha.-1-antichymotrypsin. The invention further provides analogues of .alpha.-1-antichymotrypsin that exhibit antichymotrypsin, anti-trypsin and anti-thrombin activity and methods of producing the analogues.

Abstract: Disclosed are gastric acid-resistant polymer-coated digestive enzymes/ursodeoxycholate compositions, process for their preparations and methods of treating digestive disorders, impaired liver function, cystic fibrosis, regulating the absorption of dietary cholesterol, and for dissolving gallstones by administering the compositions to a mammal in need of such treatment.