Triple-acting Lytic Enzyme Treatment of Drug-Resistant and Intracellular Staphylococcus aureus.
Multi-drug resistant bacteria are a persistent problem in modern health care, food safety and animal health. There is a need for new antimicrobials to replace over used conventional antibiotics. Here we describe engineered triple-acting staphylolytic peptidoglycan hydrolases wherein three unique antimicrobial activities from two parental proteins are combined into a single fusion protein. This effectively reduces the incidence of resistant strain development. The fusion protein reduced colonization by Staphylococcus aureus in a rat nasal colonization model, surpassing the efficacy of either parental protein. Modification of a triple-acting lytic construct with a protein transduction domain significantly enhanced both biofilm eradication and the ability to kill intracellular S. aureus as demonstrated in cultured mammary epithelial cells and in a mouse model of staphylococcal mastitis. Interestingly, the protein transduction domain was not necessary for reducing the intracellular pathogens in cultured osteoblasts or in two mouse models of osteomyelitis, highlighting the vagaries of exactly how protein transduction domains facilitate protein uptake. Bacterial cell wall degrading enzyme antimicrobials can be engineered to enhance their value as potent therapeutics.
ARS, USDA, 10300 Baltimore Ave, Beltsville, MD, USA.
Stephen C
SC
Becker
ARS, USDA, 10300 Baltimore Ave, Beltsville, MD, USA.
Dwayne R
DR
Roach
Biology, Univ. North Carolina, Charlotte, Charlotte, NC, USA.
Vinita S
VS
Chauhan
Institute for Bioscience and Biotechnology Research, University of MD, Rockville, MD, USA.|Department of Veterinary Medicine, University of MD, College Park, MD, USA.
Yang
Y
Shen
ARS, USDA, 10300 Baltimore Ave, Beltsville, MD, USA.
Juli
J
Foster-Frey
ARS, USDA, 10300 Baltimore Ave, Beltsville, MD, USA.
Anne M
AM
Powell
ARS, USDA, 10300 Baltimore Ave, Beltsville, MD, USA.
Gary
G
Bauchan
ARS, USDA, 10300 Baltimore Ave, Beltsville, MD, USA.
Richard A
RA
Lease
ARS, USDA, 10300 Baltimore Ave, Beltsville, MD, USA.
Homan
H
Mohammadi
ARS, USDA, 10300 Baltimore Ave, Beltsville, MD, USA.
William J
WJ
Harty
ARS, USDA, 10300 Baltimore Ave, Beltsville, MD, USA.
Chad
C
Simmons
ARS, USDA, 10300 Baltimore Ave, Beltsville, MD, USA.
Mathias
M
Schmelcher
ARS, USDA, 10300 Baltimore Ave, Beltsville, MD, USA.
Mary
M
Camp
Biochemistry, Univ. Alabama, Birmingham, Birmingham, AL, USA.
Shengli
S
Dong
Biochemistry, Univ. Alabama, Birmingham, Birmingham, AL, USA.
John R
JR
Baker
Biology, San Diego State University, San Diego, CA, USA.
Tamsin R
TR
Sheen
Biology, San Diego State University, San Diego, CA, USA.
Kelly S
KS
Doran
Biochemistry, Univ. Alabama, Birmingham, Birmingham, AL, USA.
David G
DG
Pritchard
University of Tennessee, Knoxville, TN, USA.
Raul A
RA
Almeida
Institute for Bioscience and Biotechnology Research, University of MD, Rockville, MD, USA.|Department of Veterinary Medicine, University of MD, College Park, MD, USA.
Daniel C
DC
Nelson
Biology, Univ. North Carolina, Charlotte, Charlotte, NC, USA.
Ian
I
Marriott
Channing Laboratory, Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA, USA.
Jean C
JC
Lee
ARS, USDA, 10300 Baltimore Ave, Beltsville, MD, USA.
David M
DM
Donovan
D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism* / Q000627:therapeutic use*; D002353:Carrier State / Q000517:prevention & control; D002478:Cells, Cultured; D004195:Disease Models, Animal; D006801:Humans; D008413:Mastitis / Q000188:drug therapy; D051379:Mice; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000235:genetics / Q000378:metabolism*; D010019:Osteomyelitis / Q000188:drug therapy; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D013203:Staphylococcal Infections / Q000188:drug therapy* / Q000517:prevention & control*; D013211:Staphylococcus aureus / Q000187:drug effects*; D016896:Treatment Outcome
D000900:Anti-Bacterial Agents; D011993:Recombinant Fusion Proteins; D009238:N-Acetylmuramoyl-L-alanine Amidase