Monofunctional glycosyltransferase of Staphylococcus aureus
AssigneeEli Lilly and Company
InventorsJo Ann Hoskins, Stanley R. Jaskunas, Jr., Robert B. Peery, Paul L. Skatrud
Life of this patent
Estimating the term…
Abstract
The invention provides isolated nucleic acid compounds encoding a novel MTG of Staphylococcus aureus. Also provided are vectors and transformed heterologous host cells for expressing the MTG and a method for identifying compounds that bind and/or inhibit the enzymatic activity of the MTG.
Claims13
An isolated monofunctional glycosyltransferase from Staphylococcus aureus having the amino acid sequence:
- Met Lys Arg Ser Asp Arg Tyr Ser Asn Ser 1 5 10 - Asn Glu His Phe Glu His Met Lys His Glu 15 20 - Pro His Tyr Asn Thr Tyr Tyr Gln Pro Val 25 30 - Gly Lys Pro Pro Lys Lys Lys Lys Ser Lys 35 40 - Arg Ile Leu Leu Lys Ile Leu Leu Thr Ile 45 50 - Leu Ile Ile Ile Ala Leu Phe Ile Gly Ile 55 60 - Met Tyr Phe Leu Ser Thr Arg Asp Asn Val 65 70 - Asp Glu Leu Arg Lys Ile Glu Asn Lys Ser 75 80 - Ser Phe Val Ser Ala Asp Asn Met Pro Glu 85 90 - Tyr Val Lys Gly Ala Phe Ile Ser Met Glu 95 100 - Asp Glu Arg Phe Tyr Asn His His Gly Phe 105 110 - Asp Leu Lys Gly Thr Thr Arg Ala Leu Phe 115 120 - Ser Thr Ile Ser Asp Arg Asp Val Gln Gly 125 130 - Gly Ser Thr Ile Thr Gln Gln Val Val Lys 135 140 - Asn Tyr Phe Tyr Asp Asn Asp Arg Ser Phe 145 150 - Thr Arg Lys Val Lys Glu Leu Phe Val Ala 155 160 - His Arg Val Glu Lys Gln Tyr Asn Lys Asn 165 170 - Glu Ile Leu Ser Phe Tyr Leu Asn Asn Ile 175 180 - Tyr Phe Gly Asp Asn Gln Tyr Thr Leu Glu 185 190 - Gly Ala Ala Asn His Tyr Phe Gly Thr Thr 195 200 - Val Asn Lys Asn Ser Thr Thr Met Ser His 205 210 - Ile Thr Val Leu Gln Ser Ala Ile Leu Ala 215 220 - Ser Lys Val Asn Ala Pro Ser Val Tyr Asn 225 230 - Ile Asn Asn Met Ser Glu Asn Phe Thr Gln 235 240 - Arg Val Ser Thr Asn Leu Glu Lys Met Lys 245 250 - Gln Gln Asn Tyr Ile Asn Glu Thr Gln Tyr 255 260 - Gln Gln Ala Met Ser Gln Leu Asn Arg 265
- which is SEQ ID NO 2.
An isolated soluble form of monofunctional glycosyltransferase from Staphylococcus aureus having the amino acid sequence which is defined by amino acid residues 68 through 269 of SEQ ID NO.2.
An isolated form of monofunctional glycosyltransferase from Staphylococcus aureus having the amino acid sequence which is defined by amino acid residues 17 through 269 of SEQ ID NO. 2.
An isolated monofunctional glycosyltransferase from Staphylococcus aureus consisting essentially of the amino acid sequence shown in SEQ ID NO. 2.
An isolated soluble form of monofunctional glycosyltransferase from Staphylococcus aureus consisting essentially of amino acid residues 68 through 269 of SEQ ID NO. 2.
An isolated form of monofunctional glycosyltransferase from Staphylococcus aureus consisting essentially of amino acid residues 17 through 269 of SEQ ID NO. 2.
A method for identifying a compound that inhibits the transglycosylase activity of a Staphylococcus aureus monofunctional glycosyltransferase, comprising the following steps:
- a) admixing:
- i) a protein selected from the group consisting of substantially pure Staphylococcus aureus monofunctional glycosyltransferase and a fragment thereof that retains transglycosylase activity,
- ii) a suitable substrate for Staphylococcus aureus monofunctional glycosyltransferase, and
- iii) a test compound to be studied for inhibitory activity against said transglycosylase activity;
- b) incubating said protein, said substrate, and said test compound together;
- c) measuring transglycosylase activity by any suitable means;
- and
- d) comparing transglycosylase activity of step c) to transglycosylase activity observed upon incubating said protein and said substrate as in step b) in the absence of said test compound.
Dependencies are inferred from the claim text.