Spectrophotometric determination of microgram quantities of protein without nucleic acid interference

Groves, William E.; Davis, Francis C.; Sells, Bruce H.

doi:10.1016/0003-2697(68)90307-2

Cited by 450 publications

(137 citation statements)

References 7 publications

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“…Contamination was relatively low for all preparations (as indicated from a mean ratio of 0·865 ± O· 024) and was within the limits suggested by Groves et al (1968) for spectrophotometric determinations of protein without nucleic acid interference.…”

Section: (A) Nucleic Acid Contaminationsupporting

confidence: 61%

“…Enzymatic activities were expressed on a protein basis using the methods of Groves et al (1968) which were described as being free from nucleic acid interference. Protein estimation by microKjeldahl determinations were made on some samples and compared with the results obtained with the Groves et al procedure using bovine serum albumin (Sigma Chemical Co.) as standard, with good agreement.…”

Section: (E) Protein Determinationsmentioning

confidence: 99%

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Enzyme Activity of the Cytochrome System of the Egg and Larva of the Cattle Tick (Boophilus Microplus)

Shanahan¹,

O'hagan²

1973

Aust. Jnl. Of Bio. Sci.

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The enzymatic activities of the succinate-<;ytochrome c reductase system, the NADH--cytochrome c reductase system, the NADH oxidase system, and cytochrome c oxidase were determined spectrophotometrically in particulate preparations of eggs and larvae of B. micro plus.All enzyme systems were detectable in the eggs of the tick and the activities remained relatively constant. At day 0, the day of hatching, an 80% increase inNADHcytochrome c reductase was recorded. No other appreciable change occurred until about day 5 when there was a sharp increase in cytochrome c oxidase activity. This was followed by an increase in succinate-<;ytochrome c reductase on day 7. NADH oxidase did not increase until about day 9.The reductases accounted for about 70% of the total oxidoreductase activity during the early larval phase but from day 9 the reductases declined, concomitant with an increase in oxidase activity. The oxidases reached a maximum of 66% of the total oxidoreductase activity about day 36.These changes in the cytochrome system are indicative of the increasing metabolic activity of the larvae with age. The increased respiratory activity also reflects the increasing demand for energy at a time when the larvae are seeking a host.Preliminary studies of the respiratory system in day 8 larvae of four strains (Yeerongpilly, Biarra, Ridgelands, and Mackay) of varying acaricide susceptibilities indicated some interstrain difference. The Mackay strain exhibited enzyme activities 2· 5-3 times greater than the Biarra strain.

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Section: (A) Nucleic Acid Contaminationsupporting

confidence: 61%

Section: (E) Protein Determinationsmentioning

confidence: 99%

Enzyme Activity of the Cytochrome System of the Egg and Larva of the Cattle Tick (Boophilus Microplus)

Shanahan¹,

O'hagan²

1973

Aust. Jnl. Of Bio. Sci.

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“…43 400 M −" :cm −" for ∆N∆C-pro-apVP, native apVP, PA protease-cleaved pro-apVP and PA protease-cleaved ∆C-proapVP. The absorption coefficient for each protein was calculated from the amino acid composition of Trp, Tyr, His, Met, Cys and cystine residues using the molar absorption coefficients 5070, 1190, 5, 4, 20 and 111, respectively [13].…”

Section: Processing Of Pro-apvp and ∆C-pro-apvp And Determination Of mentioning

confidence: 99%

Function of the N-terminal propeptide of an aminopeptidase from Vibrio proteolyticus

Zhang

Nirasawa

Nakajima

et al. 2000

Biochemical Journal

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An aminopeptidase from Vibrio proteolyticus was translated as a preproprotein consisting of four domains: a signal peptide, an N-terminal propeptide, a mature region and a C-terminal propeptide. Protein expression and analysis of the activity results demonstrated that the N-terminal propeptide was essential to the formation of the active enzyme in Escherichia coli. Urea dissolution of inclusion bodies and dialysis indicated that the N-terminal propeptide could facilitate the correct folding of the enzyme in vitro. Using l-Leu-p-nitroanilide as the substrate, the kinetic parameters (kcat and Km) of the pro-aminopeptidase and processed aminopeptidases were analysed. The results suggested that the N-terminal propeptide inhibited enzyme activity of the mature region. In contrast, the C-terminal propeptide did not show evidence of forming an active enzyme, of correctly folding in vitro or of inhibiting the active region.

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“…After the last centrifugation and resuspension, the particulate fraction and the cytosol were frozen in liquid N2 and stored at -80 ~ until the next day. Protein was measured with bovine serum albumin as standard as described by Groves et al [11].…”

Section: Preparation Of Islet Particulate and Soluble Fractionsmentioning

confidence: 99%

Characteristics of an adenylate cyclase enhancing factor from mouse pancreatic islet cytosol

1984

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Summary. The role of cytosolic components in the regulation of mouse pancreatic islet adenylate cyclase activity was studied. Addition of mouse islet cytosol (27 000 g supernatant of mouse islet sonicate), devoid of adenylate cyclase activity itself, increased adenylate cyclase activity by 93_ 17% (n =9) in the 27 000 g total particulate fraction of mouse islets. Addition of GTP stimulated adenylate cyclase activity by 91 _+ 11% (n ---13) or to the same degree as cytosol. Like GTP, the substance causing the enhancing activity of the cytosol was found to be dialysable, resistant to heat, sensitive to charcoal treatment and alkaline phosphatase and insensitive to digestion with trypsin. However, in contrast to the stimulation by GTP, the stimulation by cytosol was not inhibited by guanosine 5'-0-(2-thiodiphosphate), and furthermore, the effects of cytosol and GTP were additive. Neither NAD nor phosphoenolpyruvate stimulated adenylate cyclase activity. The cytosolic factor did not confer sensitivity towards glucose, Ca 2+ or Ca2+-calmodulin on adenylate cyclase. The results demonstrate that mouse pancreatic islets contain a phosphocompound (or several compounds) distinct from GTP and capable of markedly stimulating adenylate cyclase. The identity of the compound and its physiological significance remain to be established.

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Spectrophotometric determination of microgram quantities of protein without nucleic acid interference

Cited by 450 publications

References 7 publications

Enzyme Activity of the Cytochrome System of the Egg and Larva of the Cattle Tick (Boophilus Microplus)

Enzyme Activity of the Cytochrome System of the Egg and Larva of the Cattle Tick (Boophilus Microplus)

Function of the N-terminal propeptide of an aminopeptidase from Vibrio proteolyticus

Characteristics of an adenylate cyclase enhancing factor from mouse pancreatic islet cytosol

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