Rhipicephalus microplus serine protease inhibitor family: annotation, expression and functional characterisation assessment

Background: Rhipicephalus (Boophilus) microplus evades the host's haemostatic system through a complex protein array secreted into tick saliva. Serine protease inhibitors (serpins) conform an important component of saliva which are represented by a large protease inhibitor family in Ixodidae. These secreted and non-secreted inhibitors modulate diverse and essential proteases involved in different physiological processes. Methods: The identification of R. microplus serpin sequences was performed through a web-based bioinformatics environment called Yabi. The database search was conducted on BmiGi V1, BmiGi V2.1, five SSH libraries, Australian tick transcriptome libraries and RmiTR V1 using bioinformatics methods. Semi quantitative PCR was carried out using different adult tissues and tick development stages. The cDNA of four identified R. microplus serpins were cloned and expressed in Pichia pastoris in order to determine biological targets of these serpins utilising protease inhibition assays. Results: A total of four out of twenty-two serpins identified in our analysis are new R. microplus serpins which were named as RmS-19 to RmS-22. The analyses of DNA and predicted amino acid sequences showed high conservation of the R. microplus serpin sequences. The expression data suggested ubiquitous expression of RmS except for RmS-6 and RmS-14 that were expressed only in nymphs and adult female ovaries, respectively. RmS-19, and -20 were expressed in all tissues samples analysed showing their important role in both parasitic and non-parasitic stages of R. microplus development. RmS-21 was not detected in ovaries and RmS-22 was not identified in ovary and nymph samples but were expressed in the rest of the samples analysed. A total of four expressed recombinant serpins showed protease specific inhibition for Chymotrypsin (RmS-1 and RmS-6), Chymotrypsin / Elastase (RmS-3) and Thrombin (RmS-15). Conclusion: This study constitutes an important contribution and improvement to the knowledge about the physiologic role of R. microplus serpins during the host-tick interaction.

Importance of non-conserved distal carboxyl terminal amino acids in two peptidases belonging to the M1 family: Thermoplasma acidophilum Tricorn interacting factor F2 and Escherichia coli Peptidase N

Enzymes belonging to the M1 family play important cellular roles and the key amino acids (aa) in the catalytic domain are conserved. However, C-terminal domain aa are highly variable and demonstrate distinct differences in organization. To address a functional role for the C-terminal domain, progressive deletions were generated in Tricorn interacting factor F2 from Thermoplasma acidophilum (F2) and Peptidase N from Escherichia coli (PepN). Catalytic activity was partially reduced in PepN lacking 4 C-terminal residues (PepNΔC4) whereas it was greatly reduced in F2 lacking 10 C-terminal residues (F2ΔC10) or PepN lacking eleven C-terminal residues (PepNΔC11). Notably, expression of PepNΔC4, but not PepNΔC11, in E. coliΔpepN increased its ability to resist nutritional and high temperature stress, demonstrating physiological significance. Purified C-terminal deleted proteins demonstrated greater sensitivity to trypsin and bound stronger to 8-amino 1-napthalene sulphonic acid (ANS), revealing greater numbers of surface exposed hydrophobic aa. Also, F2 or PepN containing large aa deletions in the C-termini, but not smaller deletions, were present in high amounts in the insoluble fraction of cell extracts probably due to reduced protein solubility. Modeling studies, using the crystal structure of E. coli PepN, demonstrated increase in hydrophobic surface area and change in accessibility of several aa from buried to exposed upon deletion of C-terminal aa. Together, these studies revealed that non-conserved distal C-terminal aa repress the surface exposure of apolar aa, enhance protein solubility, and catalytic activity in two soluble and distinct members of the M1 family.

Jacobi-Schlossberg family papers, undated, 1870-2000.

The collection documents the lives Sarah Simon Jacobi (1857-1943), Freda Moritz Jacobi (1886-1939), Alice Jacobi Schlossberg (1912-1987) and Deda Schlossberg Miller (1940- ), through diaries, baby books, photographs, and correspondence. Also included in the collection are a silver card case, a silver coffee pot and creamer, and an etching of the Jacobi family home. For further information on these objects see the museum curator.

Rabb Family/Stop & Shop collection, 1912-1989.

The Rabinovitz/Rabb family arrived in Boston from Russia in the 1890s. Around 1914 they founded Economy Grocery Stores, which became Stop & Shop in 1946. In addition to building their grocery company into a successful business, the family is known for its philanthropy and active involvement in the Jewish community. The collection contains materials relating to the Rabb family and to the business operations of Stop & Shop until 1989. The materials in this collection include historical sketches, newspaper clippings, press releases, correspondence, memoranda, minutes, reports, advertisements, certificates, speeches, interviews, films, and photographs.

Israelson Family Collection

Genealogical tables for Israelson and Blum families, and a cover letter.

Strauss-Stern Family 1902-1942

Personal and official documents relating to the life of the Strauss family during the years 1902-1940: pre-marital agreement dated February 1902 and June 1904 (Ehe und Erbvertrag); partnership agreement dated on several dates in 1905; variety of Lehrvertraege, passports, documents necessary for emigration and immigration to Italy and USA. The Lehrvertrag is dated 1920, the other documents pertain to the period from 1938 to 1942. "Schulzeugnisse" from the early 1900s.

Fabian family collection 1827-1875

Three original documents: citizenship certificate and oath for Israel Fabian (June 23, 1827); certificate of family name and Prussian citizenship for Israel Fabian (December 21, 1838); appointment of widow Fabian as quartermaster (September 14, 1875).

Stillschweig Family Collection 1896-1989

Three page letter to Diane Spielman relating to resarch on Samuel Stillschweig and Jewish community in Heide.

Goldschmidt family; Hanover 1828-1842

Marriage contract; testament; Schutzbrief.

[The Rosenthal family from Münchholzhausen] 1868-1957

Vital records (originals and translations) and a family tree of the descendents of the cobbler Moses Rosenthal (1820-1896) and his wife Hannchen Stern (1815-1868) from Münchholzhausen, Germany.

Mailaender Family Collection 1962

Newspaper clippings realting to the 100th anniversary of Mailaender brewery "Bergbraeu" (1862-1962) in Fuerth, Bavaria.

My [Schoemann] family history 1811-1984

A 21 page typescript accompanied by copies of vital records and a family tree.

Jules Wertheimer family collection.

Family trees listing; newspaper clippings on Nazi period in Wertheim; inscriptions from Jewish cemetery in Schmieheim.

Beradt Family Collection 1850-1956

Documents related to Ulla Brode, née Beradt and her family.

Weinstein-Levin family trees 1862-1987

2 volumes each containing the geneaelogy of the family of Jack Weinstein: the Weinstein and the Levin families. Volumes contain photographs, newspaper clippings and other materials relevant to history of the two families.