Shellfish Tropomyosin IgE Cross-Reactivity Differs Among Edible Insect Species.

Scope: Insects are a potentially environmentally friendly alternative dietary protein source to supplement mammalian and fish sources, but potential allergenic risks are a concern. Consumption of insects may result in anaphylaxis and has been implicated in cross-reactivity with shellfish. Many allergenic proteins may be involved in cross-reactivity, including tropomyosin (TM).

P39, a novel soybean protein allergen, belongs to a plant-specific protein family and is present in protein storage vacuoles.

Soybean lecithins are seeing increasing use in industry as an emulsifier and food additive. They are also a growing source of human food allergies, which arise principally from the proteins fractionating with the lecithin fraction during manufacture. A previous study (Gu, X.

Microarray profiling of skeletal muscle sarcoplasmic reticulum proteins.

Microarrays were developed to profile the level of proteins associated with calcium regulation in sarcoplasmic reticulum (SR) isolated from porcine Longissimus muscle. The microarrays consisted of SR preparations printed onto to glass slides and probed with monoclonal antibodies to 7 target proteins. Proteins investigated included: ryanodine receptor, (RyR), dihydropyridine receptor, (DHPR), triadin (TRI), calsequestrin (CSQ), 90 kDa junctional protein (JSR90), and fast-twitch and slow-twitch SR calcium ATPases (SERCA1 and SERCA2).

C-Terminal 23 kDa polypeptide of soybean Gly m Bd 28 K is a potential allergen.

Gly m Bd 28 K is a major soybean (Glycine max Merr.) glycoprotein allergen. It was originally identified as a 28 kDa polypeptide in soybean seed flour.

Identification and analysis of a conserved immunoglobulin E-binding epitope in soybean G1a and G2a and peanut Ara h 3 glycinins.

To identify conserved immunoglobulin E (IgE)-binding epitopes among legume glycinins, we utilized recombinant soybean G2a and G2a-derived polypeptide fragments. All of these fusion polypeptides bound IgE, and the C-terminal 94-residue fragment appeared to bind more IgE. Using synthetic peptides we identified S219-N233 (S(219)GFAPEFLKEAFGVN(233)) as the dominant IgE-binding epitope.

Direct Analysis of Resveratrol in Wine by Micellar Electrokinetic Capillary Electrophoresis.

Resveratrol is an antioxidant found in grapes and other plants. It has been reported to have health benefits including anticarcinogenic activity and protection against coronary heart disease. Previous methods for its quantification in wines have relied on HPLC and GC.

Soybean glycinin G1 acidic chain shares IgE epitopes with peanut allergen Ara h 3.

Background: The identification of IgE epitopes for proteins is the first step in understanding the interaction of allergens with the immune system. Proteins from the legume family have shown in vitro cross-reactivity in IgE-binding assays, but this cross-reactivity is rarely clinically significant. Resolution of this discrepancy requires IgE epitope mapping of legume family protein allergens.

Quantitative separation of 4-hydroxyproline from skeletal muscle collagen by micellar electrokinetic capillary electrophoresis.

The phenylthiohydantoin (PTH) derivatives of 3- and 4-hydroxyproline (Hyp) were separated using micellar electrokinetic capillary electrophoresis (MEKC). The separation protocol was also used to determine Hyp content of bovine skeletal perimysial collagen preparations and whole muscle samples. Amino acids from hydrolyzed tissues were labeled using a two step procedure that involved initial reaction with o-phthalaldehyde (OPA) to modify primary amines followed by their precipitation under acidic conditions.

Capillary electrophoretic analysis of cathepsin D action on hemoglobin.

The action of purified cathepsin D on hemoglobin was examined using micellar electrokinetic chromatographic separation of peptide products. Purified cathepsin D was incubated with hemoglobin in 40 mM Na-formate pH 3.1 at 37 degrees C for varying lengths of time.

Conditions for the culture of bovine embryonic myogenic cells.

The objective of this experiment was to determine the growth characteristics of bovine embryonic muscle cells and to optimize the growth conditions for these cells using commercially-prepared media and sera. In the first study, the growth of muscle cells isolated from the hindlimb was determined by measuring DNA content. The DNA concentration was lowest (P < 0.

Determination of 3-methylhistidine by capillary electrophoresis.

The post-translational methylation of histidine to form 3-methylhistidine (3MH) is a modification principally found in contractile proteins, and thus, the level of free 3MH has been used to monitor muscle protein turnover. This work describes procedures for the capillary electrophoretic separation and determination of the phenylthiohydantoin (PTH) derivative of 3MH using uncoated fused-silica capillaries. The procedure described here utilized UV detection and resulted in a linear standard curve in the range of 2-15 pmole, which is more sensitive than previously reported HPLC methods using fluorescent detection.

Analysis of endoproteinase Arg C action on adrenocorticotrophic hormone by capillary electrophoresis and reversed-phase high-performance liquid chromatography.

The specificity and rate of cleavage of adrenocorticotrophic hormone (ACTH) peptide bonds by endoproteinase Arg C were analyzed using capillary electrophoresis (CE) and reversed-phase (C18) high-performance liquid chromatography (HPLC). Acidic cleavage products were readily resolved by CE in uncoated capillaries using low ionic strength electrolytes. However, products predicted to have a net positive charge greater than 2 or more than 4 positively charged groups per peptide did not migrate out from the capillary at low ionic strength.

Determination of native folates in milk and other dairy products by high-performance liquid chromatography.

Folates were measured in dairy products by high-performance liquid chromatography without prior sample clean-up. Detection limits for individual folates range from 0.3 to 7.

Interaction of alpha-actinin, filamin and tropomyosin with F-actin.

The abilities of alpha-actinin, filamin and tropomyosin to bind F-actin were examined by cosedimentation experiments. Results indicated that smooth muscle alpha-actinin and filamin can bind to actin filaments simultaneously with little evidence of competition. In contrast, tropomyosin exhibits marked competition with either filamin or alpha-actinin for sites on actin filaments.

A Ca2+-activated protease possibly involved in myofibrillar protein turnover. Purification from porcine muscle.

Ca2+-activated Z-disk-removing activity in the P0-40 crude muscle extracts described by Busch et al. (Busch, W. A.