Some chemical and kinetic studies of crystalline papain

Smith, Emil L.; Finkle, Bernard J.; Stockell, Anne

doi:10.1039/df9552000096

Cited by 48 publications

(16 citation statements)

References 5 publications

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“…The isolated peptides were hydrolysed in 6M HC1, 0.4% (w/v) phenol for 24 h at 110 °C (37] ; or in 12M HC1/TFA (2:1, v/v), 0.4% (w/v) phenol for 30 min at 160 °C |38} ; or in the gas phase of 6M HCI, 1% (w/v) phenol for l h at 150 °C [391 . Sometimes multiples of these (standard) times was used.…”

Section: Amino-acid Analysismentioning

confidence: 99%

The Complete Amino-Acid Sequence of the Heavy Chain of the Human Myeloma Protein WIE, an Immunoglobulin D

Friedrich

Bätge²,

Schranner³

et al. 1991

Biological Chemistry Hoppe-Seyler

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The human myeloma protein WIE is achains, O-glycosylations are confined to the hinge retype immunoglobulin D; the amino-acid sequence of gion. Furthermore, the ratios of 7V-glycosylations at its Fc part and aminoethylated heavy chain was comthe three positions are identical in IgD WAH pletely determined. The V H -part (subgroup III) be- [Takahashi, N. et al. (1984) /. Chromatogr. 317, gins W-terminally with 5-oxoproline, and it contains a 11-26.] and IgD WIE (100%, 50%, 100% ). long, unique CDR3 region. Since the constant part From the most conserved constant domain, C53, a differs from known chains by one amino-acid subthree-dimensional model was constructed to clarify stitution in the hinge region, IgD WIE probably repre-the role of its o. sp ecific substitutions and glycosylasents an allotypic variant. As in other protein t i on Die vollständige Aminosäure-Sequenz der schweren Kette des menschlichen Myelomproteins WIE, einem Immunglobulin D Zusammenfassung: Das menschliche Myelomproethylierten schweren (H-) Kette wurde vollständig ertein WIE ist ein Immunglobulin D vom -Typ; die mittelt. Der variable Teil aus der Subgruppe III beAminosäure-Sequenz vom Fc-Teil und der aminoginnt mit einem 5-Oxoproline-Rest und enthält eine Enzymes: Serine carboxypeptidase, peptidyl-L-amino-acid hydrolase (EC 3.4.16.1) (also named carboxypeptidaseY); Carboxypeptidase A, peptidyl-L-amino-acid hydrolase (EC 3.4.17.1);

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Section: Amino-acid Analysismentioning

confidence: 99%

The Complete Amino-Acid Sequence of the Heavy Chain of the Human Myeloma Protein WIE, an Immunoglobulin D

Friedrich

Bätge²,

Schranner³

et al. 1991

Biological Chemistry Hoppe-Seyler

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“…When papain or mercuripapain in 0.005 M NaHCO8 is passed through such a column, a solution of the reduced protein at its isoionic point is obtained. Columnreduced papain is active in the absence of activating agents (150) but is slowly inactivated on exposure to atmospheric oxygen. When BAL or cysteine and Versene are added to the solution the enzyme exhibits full activity and is stable in the active form.…”

Section: Activation Of Papainmentioning

confidence: 99%

The Properties of Papain

Kimmel¹,

Smith²

1957

Advances in Enzymology - And Related Areas of Molecular Biology

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“…The data were fitted to the following equation where a is the starting concentration of insulin, x is the amount of product (alanine or asparagine) formed in time t , k is the rate constant, and [El0 is the initial enzyme concentration in moles/liter using a value of 34,300 g/mole for the molecular weight of carboxypeptidase A (Smith and Stockell, 1954). The expression a -x was obtained by subtracting the moles of alanine (or asparagine) released in time t from the initial moles of insulin present.…”

mentioning

confidence: 99%

“…The initial velocity was normalized to the enxyme by dividing by the molar concentration of the enzyme to give the normalized initial velocity (v/[El0) in units of min-1. A value of 34,300 g/mole was used for the molecular weight of the carboxypeptidase A (Smith and Stockell, 1954). For each pH, the values of v/[E]o were plotted against substrate concentration ([SI).…”

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confidence: 99%

Kinetic Studies on the Action of Carboxypeptidase A on Bovine Insulin and Related Model Peptides^*

Slobin¹,

Carpenter²

1966

Biochemistry

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Several model peptides which have carboxylterminal amino acids related to those found in bovine insulin were synthesized and tested as substrates for the action of carboxypeptidase A over the pH range 6.0-9.0. The normalized initial velocities (v/[EI,) for the action of the enzyme on the carbobenzoxyglycyl derivatives of L-alanine, L-glutamine, and L-asparagine were directly proportional to the substrate concentrat ion over the substrate range 0.01-0.15 M in the pH ritnge 6.5-9.0. The second-order rate constants ( k ) for these substrates exhibited maximum values at pH 7.0-7.5. Carbobenzoxyglycyl derivatives of L-glutamic acid and Laspartic acid were much poorer substrates than the corresponding amides. The initial velocities for the action of carboxypeptidase A on these substrates had maximum values at pH 6.0 (or below) and fell off rapidly at pH values above 7.0. The difference in the pH optimum between the neutral and acidic: amino acid residues explains in part the differences previously encountered in the pH profile for the carboxypeptidasecatalyzed hydrolysis of asparagine or aspartic acid from insulin and desamido-insulin, respectively. In studies on insulin the rate of release of carboxyl-terminal alanine (from the B chain) and of asparagine (from the A chain) was markedly increased as the concentration of the insulin was lowered from 10 mg/rril to 1 mg/ml C arboxypeptidase A has been used extensively as a tool for elucidating the primary structurl: of proteins, and much valuable information about its mode of attack on proteins has been accumulated i: Davie et at., 1959;Neurath, 1960). The present investigation stems from earlier work in this laboratory in which carboxypeptidase A was used to determine the difference in the primary structure between insulin and desamido-insulin (Slobin and Carpenter, 1963a,b). We were attracted to a study of the kinetics of the insulin-carboxypeptidase A system by several features which made it appear suitable Congress of Biochemistry (1964). Abstracted from a thesis submitted by Lawrence I. Slobin in partial fulfillmerit of the requirements of the Ph.D. degree at the University of California, Berkelev. 1964. in the pH range 7.5-9.0. The rate of release of both amino acids increased in going from pH 7.5 to 9.0 at all concentrations of insulin. Furthermore, increasing the salt concentration brought about a decrease in the rate of release of carboxyl-terminal amino acids from insulin. These results on insulin are in contrast to those predicted from studies of the action of carboxypeptidase on model peptides. However, there is a good correlation bet ween conditions which bring about depolymerization of insulin (as judged by ultracentrifugal studies) and those which increase the rate of attack of carboxypeptidase on insulin. This correlation suggests that the action of carboxypeptidase on insulin is largely dependent on the degree of polymerization of the insulin.Rotatory dispersion studies have been made of the products of digestion : desalanine-insulin and desalanine-des...

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Some chemical and kinetic studies of crystalline papain

Cited by 48 publications

References 5 publications

The Complete Amino-Acid Sequence of the Heavy Chain of the Human Myeloma Protein WIE, an Immunoglobulin D

The Complete Amino-Acid Sequence of the Heavy Chain of the Human Myeloma Protein WIE, an Immunoglobulin D

The Properties of Papain

Kinetic Studies on the Action of Carboxypeptidase A on Bovine Insulin and Related Model Peptides^*

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Some chemical and kinetic studies of crystalline papain

Cited by 48 publications

References 5 publications

The Complete Amino-Acid Sequence of the Heavy Chain of the Human Myeloma Protein WIE, an Immunoglobulin D

The Complete Amino-Acid Sequence of the Heavy Chain of the Human Myeloma Protein WIE, an Immunoglobulin D

The Properties of Papain

Kinetic Studies on the Action of Carboxypeptidase A on Bovine Insulin and Related Model Peptides*

Contact Info

Product

Resources

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Kinetic Studies on the Action of Carboxypeptidase A on Bovine Insulin and Related Model Peptides^*