Amino Acid Analysis of Submicrogram Hydrolyzate Samples

Bidlingmeyer, Brian A.; Tarvin, Thomas L.; Cohen, Steven A.

doi:10.1007/978-1-59259-480-1_15

Cited by 19 publications

(16 citation statements)

References 7 publications

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“…The hydrolysis of ATP, expressed as ATP/s per myosin head, was calculated by considering the molecular mass of native myosin-V dimers to be 640 kDa (7). The final conditions used in the assays were as follows: K-EDTA-ATPase, 20 (18), using the Schwartzenbach dissociation constants for Ca-EGTA. The dependence of Mg-ATPase activity on actin concentration was determined by addition of F-actin over a range of concentrations from 0.1-60 M. Phalloidin (1:1 molar ratio with actin) was added to solutions containing 0.1 to 10 M actin to stabilize the actin as filaments.…”

Section: Methodsmentioning

confidence: 99%

Enzymatic Characterization and Functional Domain Mapping of Brain Myosin-V

Nascimento

Cheney

Tauhata

et al. 1996

Journal of Biological Chemistry

120

131

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Although the conventional myosins (myosins-II) responsible for processes such as muscle contraction and cytokinesis have been intensively studied, relatively little is known about the properties of the unconventional myosins. One widely distributed group of unconventional myosins, the class V myosins, have been suggested to play a role in organelle transport or membrane targeting (reviewed in Refs. 1 and 2). Brain myosin-V (BM-V 1 ), which was originally identified as a calmodulin (CaM)-binding protein in vertebrate brain, is the only member of this class of unconventional myosins to be purified and characterized biochemically (3-7). The class V myosins share a common domain structure consisting of an N-terminal head domain containing the actin-binding and ATP hydrolysis sites, an extended neck domain containing six IQ motifs (which form binding sites for CaM and/or related light chains), and a tail domain consisting of a region predicted to form coiled coils attached to a globular region of unknown function (7,8). The hypothetical functions of the class V myosins and their novel domain structure, particularly the extended neck domain, raise the obvious question of how the basic biochemical properties of the class V myosins compare with those of other types of myosins.Among the critically important properties of a molecular motor are its steady-state ATPase activity and the factors which regulate this activity. Known myosins are regulated either indirectly via actin-associated proteins such as troponin/ tropomyosin (as in vertebrate skeletal muscle myosin) or directly via the subunits of the myosin molecule itself. In the latter type of direct "myosin-linked" regulation, the light chains associated with the neck domain often function as the regulating subunits. The myosin light chains are all members of the CaM superfamily of proteins, although not all of them have retained the ability to bind to Ca 2ϩ . Vertebrate non-muscle and smooth muscle myosins-II are "turned on" by phosphorylation of their regulatory light chains by Ca 2ϩ /CaM-dependent myosin light chain kinase (9), whereas molluscan myosin-II and vertebrate brush border (BB) myosin-I are regulated by the direct binding of Ca 2ϩ to their light chains (10, 11). Like BB myosin-I, BM-V has multiple CaM light chains that remain bound in the absence of Ca 2ϩ (3,5,7). The neck domain of BM-V has been shown to be the precise site of CaM binding (8), suggesting that this myosin might also be directly regulated by Ca 2ϩ binding. The initial reports of BM-V ATPase activity also indicated that this protein is activated by Ca 2ϩ (5, 7), although the K ATPase for Ca 2ϩ and F-actin were not determined. The tail domains of myosins are unique to each class in the myosin superfamily and probably reflect the specific functions

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Section: Methodsmentioning

confidence: 99%

Enzymatic Characterization and Functional Domain Mapping of Brain Myosin-V

Nascimento

Cheney

Tauhata

et al. 1996

Journal of Biological Chemistry

120

131

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“…A fixed amount (45 nmol) of the internal standard L-citrulline (Aldrich, Steinheim, Germany) was added to aliquots of the dissolved hydrolysates, and derivatives of the amino acids were made with phenylisothiocyanate (Pierce, Rockford, IL) using a standard procedure. 15 The deposition of hydroxyproline (hpro) (nmol/mg) and proline (pro) (nmol/mg) in the ePTFE tubes implanted in the upper arm were measured by HPLC. The ratio of hpro to pro was calculated from an average obtained in each of the two tubes.…”

Section: High-performance Liquid Chromatography (Hplc) For Hydroxypromentioning

confidence: 99%

The Effect of Preoperative Radiotherapy on Systemic Collagen Deposition and Postoperative Infective Complications in Rectal Cancer Patients

Johnson

Jørgensen

Adawi

et al. 2005

Diseases of the Colon &Amp; Rectum

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“…The plasma amino acid profiles, including glutamate, arginine, proline, ornithine, and citrulline, were analyzed using reversed-phase HPLC. 20) All samples for the ELISA and colorimetric assays were analyzed in one assay batch in duplicate. The inter-assay coefficients of variance were 5-10%.…”

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confidence: 99%

Dose Effects of Chronically Infused Nitric Oxide Synthase Inhibitor NG-Nitro-L-arginine Methyl Ester on Anabolic Response and Arginine Metabolism in Rats with Subacute Peritonitis

Hsiao

Lee

Tsao

et al. 2011

Biological & Pharmaceutical Bulletin

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Patients with peritonitis develop arginine deficiency due to decreased consumption, impaired absorption, and increased utilization of arginine.1,2) Arginine, a precursor for the synthesis of nitric oxide (NO), urea, creatinine, glutamate, proline, and polyamines, is a conditionally essential amino acid in stressed conditions, especially in inflammation. 1,3,4) Green et al. 5) was the first research group to show that nitrate/nitrite concentrations and the severity of infection are closely correlated. Subsequent studies showed that patients with peritonitis have uncontrolled activation of inducible nitric oxide synthase (NOS), which results in NO overproduction and the following sepsis.6) Therefore, inhibition of NOS might be a useful strategy to treat arginine deficiency and inhibit excess NO production. 6,7) In animal and human studies, the results of NOS inhibition in inflammation have been ambiguous. For example, N G -nitro-L-arginine methyl ester (L-NAME), a nonselective NOS inhibitor which inhibits both constitutive and inducible NOS, effectively ameliorated the inflammatory lesions in the skin of zinc-deficient rats, 8) improved the myocardial function in sepsis, 9) and attenuated the lipopolysaccharide-induced peritoneal permeability and NO release in mice.10) On the contrary, some studies have shown that L-NAME may cause hypertension, augment the expressions of interferon (IFN)-g and interleukin (IL)-2, and result in markedly severe disease in rats with T cell-dependent autoimmune interstitial nephritis.11) The adverse effects of NOS inhibitors have also been observed in subjects with hemodynamic instability and sepsis. [12][13][14] Evidence suggests that the therapeutic effects of NOS inhibitors may depend on the dose, timing, and the severity and category of diseases.In clinical practice, a number of in hospital patients with subacute peritonitis developed chronic malnutrition and inflammation. Administration of total parenteral nutrition solution is an efficient method to supply all daily nutrition requirements; however, it is associated with occurrence of increased complicated infections.15) Zheng et al. 16) indicated that long-term parenteral nutrition-induced liver steatosis may be attenuated by L-NAME administration. Up to now, only few reports have described the long-term application of L-NAME for subacute peritonitis. Additionally, no study has ever addressed the effects of L-NAME on the inflammatory response and amino acid metabolism in subjects administered parenteral nutrition. In our study, we used a rat model with subacute peritonitis induced by cecal puncture and administered total parenteral nutrition solution to provide adequate nutrition, and we investigated the dose effects of parenterally supplemented L-NAME on the anabolism, inflammatory response, and arginine-derived metabolites such as the intermediates of the urea cycle, circulating profile of amino acids, NO, and cytokines. MATERIALS AND METHODS Animals and Experimental DesignMale Wistar rats (8-9 weeks old), initially weighi...

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Amino Acid Analysis of Submicrogram Hydrolyzate Samples

Cited by 19 publications

References 7 publications

Enzymatic Characterization and Functional Domain Mapping of Brain Myosin-V

Enzymatic Characterization and Functional Domain Mapping of Brain Myosin-V

The Effect of Preoperative Radiotherapy on Systemic Collagen Deposition and Postoperative Infective Complications in Rectal Cancer Patients

Dose Effects of Chronically Infused Nitric Oxide Synthase Inhibitor NG-Nitro-L-arginine Methyl Ester on Anabolic Response and Arginine Metabolism in Rats with Subacute Peritonitis

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