The Catharanthus roseus 16-hydroxytabersonine O-methyltransferase involved in vindoline biosynthesis /

Madagascar periwinkle (Catharanthus roseus) produces the well known and remarkably complex dimeric anticancer alkaloids vinblastine and vincristine that are derived by coupling vindoline and catharanthine monomers. This thesis describes the novel application of carborundum abrasion (CA) technique as a tool for large scale isolation of leaf epidermis enriched proteins. This technique was used to facilitate the purification to apparent homogeneity of 16-hydroxytabersonine-16-0-methyltransferse (l60MT) that catalyses the second step in the 6 step pathway that converts tabersonine into vindoline. This versatile tool was also used to harvest leaf epidermis enriched mRNAs that facilitated the molecular cloning of the 160MT. Functional expression and biochemical characterization of recombinant 160MT enzyme showed that it had a very narrow substrate specificity and high affinity for 16-hydroxytabersonine, since other closely related monoterpene indole alkaloids (MIAs) did not act as substrates. In addition to allowing the cloning of this gene, CA technique clearly showed that 160MT is predominantly expressed in Catharanthus leaf epidermis, in contrast to several other OMTs that appear to be expressed in other Catharanthus tissues. The results provide compelling evidence that most of the pathway for vindoline biosynthesis including the 0- methylation of 16-hydroxytabersonine occurs exclusively in leaf epidermis, with subsequent steps occurring in other leaf cell types. Small molecule O-methyltransferases (OMTs) (E.C. 2.1.1.6.x) catalyze the transfer of the reactive methyl group of S-adenosyl-L-methionine (SAM) to free hydroxyl groups of acceptor molecules. Plant OMTs, unlike their monomeric mammalian homologues, exist as functional homodimers. While the biological advantages for dimer fonnation with plant OMTs remain to be established, studies with OMTs from the benzylisoquinoline producing plant, Thalictrum tuberosum, showed that co-expression of 2 recombinant OMTs produced novel substrate specificities not found when each rOMT was expressed individually (Frick, Kutchan, 1999) . These results suggest that OMTs can fonn heterodimers that confer novel substrate specificities not possible with the homodimer alone. The present study describes a 160MT model based strategy attempting to modify the substrate specificity by site-specific mutagenesis. Our failure to generate altered substrate acceptance profiles in our 160MT mutants has lead us to study the biochemical properties ofhomodimers and heterodimers. Experimental evidence is provided to show that active sites found on OMT dimers function independently and that bifunctional heterodimeric OMTs may be fonned in vivo to produce a broader and more diverse range of natural products in plants.

Chapman family home, Fullerton, California, 1908

Chapman family residence, Fullerton, California, ca. 1908. Mounted on heavy weight card stock with small calendar for 1908. "Miss Ethel Chapman" written in ink on top of calendar.

Annual report of the Commissioners of the State Reservation at Niagara, 1907-1908.

Report year irregular.

Annual report of the Commissioners of the State Reservation at Niagara, 1908-1909.

Report year irregular.

Niagara Parks Commission annual report, 1908.

Printed by Order of the Legislative Assembly of Ontario.

Niagara-Welland Power Company Limited fonds, 1833, 1852, 1904-1906, 1909, n.d.

The Welland Power and Supply Canal Company Limited, established in 1893 and incorporated in 1894 with a capital stock of $500,000. The aim of the company was to harness the natural water supply of the Niagara and Welland Rivers. In 1898 the Canadian Electrical News published a report by Henry Symons, QC outlining the main project of the company. This project involves the construction of a canal from the Welland River to the brow of the mountain at Thorold, a distance of 8 miles; the construction at Thorold of a power house, and from Thorold to Lake Ontario, a raceway by which to carry water into the lake. The estimate for the machinery to generate 100,000 horse power is £125,000; for transmission line to Toronto at a voltage of 10,000….The total estimate therefore amounts to £2,452,162, or roughly speaking, $12,000,000. Source: Canadian Electrical News, August 1898, p. 172. In 1899 the company officers petitioned the federal government desiring a name change to the Niagara-Welland Power Company Limited. Officers of the company were Harry Symons, President; Charles A. Hesson, Vice-President; and M.R. O’Loughlin, James B. Sheehan, James S. Haydon, Frederick K. Foster, directors; John S. Campbell, secretary-treasurer. The company’s head offices were located in St. Catharines, with a New York (City) office on Broad Street. In 1905 and 1909 the company petitioned the federal government for additional time to construct its works, which was granted. The company had until May 16, 1915 to complete construction. John S. Campbell (1860-1950) was a graduate of the University of Toronto and Osgoode Hall. During his university years John began his military career first in "K" Company, Queens Own rifles and then later as Commanding Officer of the 19th Lincoln Regiment, from 1906 to 1910. Upon his return to St. Catharines John Campbell served as secretary in the St. Catharines Garrison Club, a social club for military men begun in 1899. After being called to the Bar, he became a partner in the firm of Campbell and McCarron and was appointed to the bench in 1916, serving until retirement in 1934. Judge Campbell served as an alderman for several terms and was the mayor of St. Catharines in 1908 and 1909. He also served as the first chairman of the St. Catharines Public Utilities in 1914. John S. Campbell was married to Elizabeth Oille, daughter of Jerome B. and Charlotte (St. John) Oille. The family home "Cruachan" was located at 32 Church St.

F. R. Lalor and Wm. Jaques Papers, 1910

F.R. [Francis Ramsey] Lalor (1856-1929) was born in St. Catharines, Ont. but lived most of his life in Dunnville, Haldimand County, Ont. He was a merchant and manufacturer. In 1903 he and a few partners founded the Monarch Knitting Co. Lalor was also an exporter of hardwood ashes used for agricultural purposes as fertilizer. Lalor was active in politics, he was a Conservative and the member of parliament for Haldimand, having been elected in the 1904, 1908, 1911 and 1917 elections. William Jaques lived in Simcoe, Norfolk County, Ontario. He was a junk dealer by profession.

Proceedings : Grand Lodge, A.F. & A.M. of Canada in the Province of Ontario - 1908

On spine : The Grand Lodge of Canada in the Province of Ontario.

Second Welland Canal - Book 2, Survey Map 5 - Locks 16, 17 and 18 in Grantham

Survey map of the Second Welland Canal created by the Welland Canal Company showing the Grantham Township at Merritton. Identified structures associated with the Canal include Locks 15, 16, 17, and 18, and the towing path. The surveyors' measurements and notes can be seen in red and black ink and pencil. Local area landmarks are also identified and include streets and roads (ex. Hartzel Road and Macadamized Road), Lybster Cotton Mill, St. Catharines Paper Mill, J. Brown Cotton Mills, hydraulic race, a pond, several barns, and a number of structures (likely houses or cabins) belonging to: J. McNamara, M. Moran, A. Delany, T. Joyce, J. Delany, C. Blake, F. Weaver, W. Leeson, and Mrs. Aikins. Properties of note are: Concession 10 Lots 11 and 12. A number of reserved properties exist and are outlined in blue. They include three reserved properties for lock lots, and one large property reserved for a quarry.

Second Welland Canal - Book 2, Survey Map 16 - Deep Cut in Thorold

Survey map of the Second Welland Canal created by the Welland Canal Company showing the canal in the Thorold Township between Allanburg and Port Robinson. Identified structures and features associated with the Canal include the Deep Cut and the towing path. The surveyors' measurements and notes can be seen in red and black ink and pencil. Local area landmarks are also identified and include streets and roads (ex. Road to Port Allanburg), and the Spoil Bank. Properties and property owners of note are: Lots 185, 186, and 187, J. J. Church and H. Vanderburgh. Four properties adjacent to the canal are outlined in blue and labeled J through M, with L and K belonging to John Beatty, M belonging to John Coulter, and J belonging to G. Jordan (formerly belonging to John Coleman Jordan).