Cooperativity and switching within the three-state model of muscle regulation.

Thin filament regulation is mediated by the presence of tropomyosin (Tm) and troponin (Tn) on the actin filament. Binding of Tm alone induces two states, closed and open (with the equilibrium between them defined by KT), which differ in their affinity for myosin subfragment 1 (S1). Cooperative switching between the states results in characteristic sigmoidal myosin S1 binding curves.

Absence of 185delAG mutation of the BRCA1 gene and 6174delT mutation of the BRCA2 gene in Ashkenazi Jewish men with prostate cancer.

Epidemiological studies have demonstrated a clustering of breast and prostate cancers in some families. Moreover, there is an increase in the number of cases of prostate cancer in families with inherited mutations of the breast cancer susceptibility gene BRCA1. We assessed the role of BRCA1 and BRCA2 in prostate cancer.

The stability of tropomyosin at acid pH: effects of anion binding.

The alpha-helical coiled-coil tropomyosin homodimer, alpha alphaTm, unfolds cooperatively with T1/2 = 47 degreesC, at neutral pH, 0.5 M NaCl. At pH 2, where each chain contains 55 positive charges, no cooperative unfolding occurs to at least 80 degreesC.

The muscle thin filament as a classical cooperative/allosteric regulatory system.

It is generally accepted that the regulation of muscle contraction involves cooperative and allosteric interactions among the protein components, actin, myosin, tropomyosin and troponin. But, as yet, the individual role of each component has not been clearly identified. Here we compare the properties of the components of the muscle regulatory system with the corresponding components of two systems, hemoglobin and aspartate transcarbamylase, that are well described by the classical Monod, Wyman and Changeux (MWC) model.

Unfolding domains and tryptophan accessibility of a 59 kDa coiled-coil light meromyosin.

Light meromyosin (LMM 77), the C-terminal proteolytic peptide from myosin rod, is a 900 A coiled-coil that contains two pairs of tryptophan residues in d-positions of the heptad repeat (abcdefg)n. Previous studies showed that LMM 77 unfolded in two transitions and suggested that both Trp pairs were located in the least stable unfolding domain. Here, the thermal and denaturant unfolding properties of LMM 59, a recombinant N-terminal truncated LMM, containing only one of the Trp pairs, was compared to LMM 77.

Actin-tropomyosin activation of myosin subfragment 1 ATPase and thin filament cooperativity. The role of tropomyosin flexibility and end-to-end interactions.

Tropomyosin (Tm) bound to actin induces cooperative activation of actomyosin subfragment 1 (actin-S1) ATPase, observed as a sigmoid ATPase vs [S1] dependence. The activation is much steeper for gizzard muscle Tm (GTm) than for rabbit skeletal Tm (RSTm). To investigate if this greater cooperativity is due to increased communication between GTms along the thin filament, we studied effects of S1 binding on the state of actin-Tm using the fluorescence of pyrene-labeled Tm.

Comparison of pediatric and adult IgE antibody binding to fish proteins.

Background: Allergic reactions to fish are a common cause of food allergy.

Objective: We compared the binding of pediatric and adult fish-allergic patient IgE antibodies to fish proteins.

Methods: Clinical histories of fish allergy were confirmed by prick skin tests, RAST and if possible, with blinded oral food challenges.

Quantification of the major brown shrimp allergen Pen a 1 (tropomyosin) by a monoclonal antibody-based sandwich ELISA.

Background: Among 13 allergens found in extracts of cooked brown shrimp (Penaeus aztecus) the 36 kd muscle protein tropomyosin has been identified as the only major shrimp allergen (Pen a 1). Cross-reacting molecules with similar molecular weights were detected in other crustacea species such as crab, lobster, and crawfish. Because Pen a 1 and Pen a 1-like allergens are important in crustacea allergy, the aim of this study was to develop a monoclonal antibody (mAb)-based sandwich ELISA to quantify Pen a 1 and to evaluate Pen a 1 levels in four commercial shrimp, crab, and lobster extracts.

Characterization of recombinant shrimp allergen Pen a 1 (tropomyosin).

Tropomyosin (Pen a 1) from brown shrimp, Penaeus aztecus, has been identified as the only major shrimp allergen. Since beef, pork and chicken are other tropomyosin-containing foods that are not very allergenic, tropomyosins can serve to investigate the contribution of the structural properties of a protein to its allergenicity. The aim of this study was to determine the primary structure of Pen a 1 and to identify IgE-binding epitopes.

Recombinant proteins in newly developed foods: identification of allergenic activity.

A number of agricultural crops are being modified for various purposes using recombinant DNA technology. Since transferred genes may code for proteins that are ordinarily not present, there is concern about the potential allergenicity of these new varieties. The safety evaluation of transgenic foods is relatively easy when the allergenicity of the gene source is known.

Effects of two familial hypertrophic cardiomyopathy-causing mutations on alpha-tropomyosin structure and function.

Missense mutations in alpha-tropomyosin can cause familial hypertrophic cardiomyopathy. The effects of two of these, Asp175Asn and Glu180Gly, have been tested on the structure and function of recombinant human tropomyosin expressed in Escherichia coli. The F-actin affinity (measured by cosedimentation) of Glu180Gly was similar to that of wild-type, but Asp175Asn was more than 2-fold weaker, whether or not troponin was present.

Detection of snow-crab antigens by air sampling of a snow-crab production plant.

Background And Objective: We previously assessed the prevalence of occupational asthma (OA) to snow-crab in production plant workers. We also showed that this type of OA is related to immediate immunological reactivity as demonstrated by skin reactivity and increased specific IgE antibodies. However, we did not show that snow-crab antigens causing immunological reactivity and OA could be found in the air sampling in the plants atmosphere.

Identification of the first major allergen of a squid (Todarodes pacificus).

Background: In Japan, squid is an important seafood, and some patients with food allergies are sensitive to squid. There has been no report, however, describing the major allergens of squid.

Objective: To characterize squid allergens, we isolated a major allergen from the Pacific flying squid (Todarodes pacificus) and compared it with a major allergen from a shrimp (Penaeus orientalis).

Occupational asthma caused by aromatic herbs.

We describe a subject with occupational asthma caused by several aromatic herbs: thyme, rosemary, bay leaf, and garlic. The diagnosis was confirmed by several inhalation challenges in the laboratory. Although immediate skin reactivity was demonstrated to the herbs the subject reacted to by inhalation, RAST showed that garlic was the most potent allergen by weight, the other herbs showing less reactivity.

Reverse transcriptase-polymerase chain reaction for prostate-specific antigen may be a prognostic indicator in breast cancer.

Among women with node-negative breast cancer and small tumours, it is important to identify those with tumours that will recur, so that they may receive adjuvant therapy, while sparing those with tumours that will not recur the hazards of adjuvant treatment. A reverse transcriptase-polymerase chain reaction (RT-PCR) for prostate-specific antigen (PSA) may be used to identify circulating metastatic cells in patients with prostate cancer. Approximately 30% of breast cancer cells also produce PSA.

Allergy to cockroaches in patients with asthma and rhinitis in an urban area (Madrid).

Previous studies have established that cockroach allergens are important sensitizing agents in the induction of rhinitis and asthma principally in urban areas. This study was undertaken to assess skin test reactivity and specific IgE antibody reactivity to extracts of Periplaneta americana (P.a.