The influence of water on the glucose affinity of hexokinase

In the present thesis, the role of hydration during the glucose induced conformational change of hexokinase is investigated. This is accomplished by applying the osmotic stress technique. The osmotic stress technique is founded on varying of the activity of water in a system in order to determine ifs effects. This is accomplished by adding inert solute molecules that are excluded from the system under study. The solute molecules used within the present investigation are Polyethylene glycols (PEGs). PEGs aid in the removal of water from hexokinase by exerting osmotic pressure. The osmotic pressures of the PEG solutions are also measured with both vapour pressure osmometry and secondary osmometry with phospholipids. An interesting discovery is made in that the osmotic pressures of PEG and co-solute solutions are non-additive. This indicates that PEG concentrates co-solutes in solution by making a certain proportion of the water inaccessible. Glucose binding was measured fluorometrically and the glucose equilibrium dissociation constant (GEDC) of hexokinase is measured in solutions containing the different MW PEGs. Changes in the sensitivity of the glucose affinity with osmotic pressure allows the calculation of the change in the numbers of polymer-inaccessible water molecules upon the binding of glucose to hexokinase ~Nw. It was determined the ~Nw decreases with increases in osmotic pressure in the presence of all MW PEGs. ~Nw decreases from values between 45-290 water molecules at low pressure to approximately 15 at high pressure. There is also a molecular weight dependence observed. There are large decreases in ~Nw with osmotic pressure in the presence of PEGs above MW 1000. However, below MW 1500 changes in ~Nw with osmotic pressure are relatively small. These findings are interpreted with respect to two possible mechanisms involving changes in the conformation of hexokinase u~der osmotic pressure and the access of the PEG molecules to water surrounding hexokinase.

Barnes Wines: Canada’s Oldest Winery flyer. n.d.

In 1873 George Barnes, Andrew Skinner, James Skinner, John Young Reid, Charles Robert Murray, George Magan, Thomas Barnes and Robert Duncan applied for, and received a charter for a commercial winery which would be called The Ontario Grape Growing and Wine Manufacturing Company Limited. It opened in 1894 and became known as Barnes Wines Limited. In 1973 the company completed a merger with Reckitt and Coleman (Canada) Limited. The winery operated until 1988 and was located on the banks of the old Welland Canal in St. Catharines, Ontario. The company produced a complete line of table wines, dessert wines, sherries, ports, and both crackling and sparkling wines. Barnes Wines called itself “Canada’s oldest winery” at the time of the printing of this flyer.

Barnes Wines Fonds

4 Minute Books dated: 1880, 1892, 1901, 1934, 1948-1950, 1952-1953, 1955, 1958, 1962 – 1975, 1981, 1984, 1986; 3 Common stock books dated: 1934-1935, 1937-1941, 1946, 1948-1950, 1955-1956, 1958, 1961-1974, 1981 and 2 Class A stock books dated:1948 - 1951

By-Law no. 12

A by-law begins: "It was then moved by R.S. Kinner and Seconded by George Barnes that by Law No. 12 be read. Whereas by Identure bearing date thirteenth day of August A.D. 1873 made between one George Barnes of the first part thereof and Andrew S. Kinner and Charles Robert Murray of the second part thereof which said Identure was duly registered in the Registry Office for Lands for the said County of Lincoln on the 14th day of August A.D. 1880 in Book 4 for the Township of Louth as No. 1534 the said George Barnes granted and conveyed the Lands and premises therein described to the said Andrew S. Kinner and Charles Robert Murray and their heirs and assigns as joint tenants and not as tenants as common upon the trusts and for the purposes therein expressed. And Whereas the said Andrew S. Kinner died on about the 13th day of June 1877 A.D. without having made any appointment under the provisions of the said in part recited Identure.."

Photograph - Robert Houston

A Photograph of Robert Houston of Port Dalhousie. He died 6 December 1889 at age 44 years and is buried in the Anglican Cemetary of Port Dalhousie. The photograph was taken at Thos. Charles, Photographer on Ontario Street in St. Catharines.

Photograph - Robert Stanfield and Sean O'Sullivan, 1972

A photograph of Robert Stanfield with Sean O'Sullivan in his office, House of Commons, June 1972.