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  • 1
    ISSN: 1573-5001
    Keywords: Villin 14T ; Heteronuclear multidimensional NMR ; Chemical shift assignments ; Hydrogen exchange ; Calcium titration ; pH titration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Sequence-specific assignments have been made for the 1H, 15N, 13C and 13CO resonances of 14T, the 126-residue amino-terminal domain of the actin-severing protein villin. Villin is a member of a family of proteins that regulate cytoskeletal actin by severing, capping and nucleating actin filaments. Actin binding is dependent on calcium and disrupted by phosphatidyl inositol 4,5-bisphosphate. Actin-severing proteins are built from three or six repeats of a conserved domain, represented by 14T. Expression in Escherichia coli facilitated incorporation of 15N and 13C isotopes and application of triple-resonance, backbone-directed strategies for the sequential assignments. Elements of regular secondary structure have been identified by characteristic patterns of NOE cross peaks and values of vicinal 3JH n Hα coupling constants. Amide protons that exchange slowly (rates less than 1.0×10-4 per min) are concentrated in the central β-sheet and the second and third α-helices, suggesting that these elements of secondary structure form very stable hydrogen bonds. Assignments for the amide nitrogens and protons have been examined as a function of pH and calcium concentration. Based on the conservation of chemical shifts in the core of the domain, villin 14T maintains the same overall fold in the pH range from 4.15 to 6.91 and the calcium range from 0 to 50 mM. The calcium data indicate the presence of two calcium-binding sites and suggest their locations.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Developmental Dynamics 203 (1995), S. 141-151 
    ISSN: 1058-8388
    Keywords: Fimbrin ; Plastin ; Intestinal epithelium ; Visceral endoderm ; Microvilli ; Adhesion ; Actin ; Development ; Embryogenesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: During the differentiation of the intestine epithelium, three cytoskeletal proteins, villin, fimbrin, and myosin I, are sequentially expressed and localized to the apical membrane. Recently, we found that in the adult mouse and human, three fimbrin isoforms are expressed in a cell specific manner. I-fimbrin is expressed by intestine and kidney epithelial cells, L-fimbrin is expressed by leukocytes and many tumors, while T-fimbrin is expressed by various cells and tissues. Because non-intestinal isoforms of fimbrin could be expressed early in development, the expression of fimbrin isoforms during days 10.5 to 16.5 of intestine development was investigated. By immunofluorescence microscopy, T-fimbrin was detected in the early stages of intestinal epithelial cell differentiation until day 14.5 and was localized predominantly at the apical surface. L-fimbrin was also detected during this period but it was localized to the basal surface of the epithelium instead of the apical surface. By day 16.5 no L or T-fimbrin was detected in the epithelium. I-fimbrin was first detected at day 14.5 and a brush border-like apical localization pattern was seen by day 16.5. Unlike the intestinal cells, the visceral endoderm expressed I, L, and T-fimbrin throughout the period examined, with the level of I-fimbrin increasing as time progresses. L-fimbrin was more evident at the earlier stage than at the later stage of the development. Collectively, these results suggest that three fimbrin isoforms play different roles during epithelial cell differentiation. T- and I-fimbrin expression could be critical for the formation and extension of the microvilli whereas L-fimbrin may play a role in controlling cell adhesion. ©1995 Wiley-Liss, Inc.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
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