The cell surface protein MUL_3720 confers binding of the skin pathogen Mycobacterium ulcerans to sulfated glycans and keratin

Abstract

Mycobacterium ulcerans is the causative agent of the chronic, necrotizing skin disease Buruli ulcer. Modes of transmission and molecular mechanisms involved in the establishment of M. ulcerans infections are poorly understood. Interactions with host glycans are often crucial in bacterial pathogenesis and the 22 kDa M. ulcerans protein MUL_3720 has a putative role in host cell attachment. It has a predicted N-terminal lectin domain and a C-terminal peptidoglycan-binding domain and is highly expressed on the surface of the bacilli. Here we report the glycan-binding repertoire of whole, fixed M. ulcerans bacteria and of purified, recombinant MUL_3720. On an array comprising 368 diverse biologically relevant glycan structures, M. ulcerans cells showed binding to 64 glycan structures, representing several distinct classes of glycans, including sulfated structures. MUL_3720 bound only to glycans containing sulfated galactose and GalNAc, such as glycans known to be associated with keratins isolated from human skin. Surface plasmon resonance studies demonstrated that both whole, fixed M. ulcerans cells and MUL_3720 show high affinity interactions with both glycans and human skin keratin extracts. This MUL_3720-mediated interaction with glycans associated with human skin keratin may contribute to the pathobiology of Buruli ulcer. Mycobacterium ulcerans causes a skin-based disease known as Buruli ulcer. How the bacteria are transmitted and what mechanisms they use to establish the infection of the skin is poorly understood. The only well characterized bacterial factor in Buruli ulcer pathogenesis is mycolactone, a toxin produced by the bacteria. Mycolactone causes apoptosis in human cells, leading to destruction of the skin around extracellular clusters of the mycobacteria. Human cells, like cells of all orders of life, are coated in complex sugar structures and these glycans are one of the major targets of bacteria and viruses for the interaction with host cells. Here we describe the glycan binding of whole Mycobacterium ulcerans cells and a mycobacterial protein, MUL_3720, thought to be involved in glycan binding. We show that both the bacterial cells and MUL_3720 bind to glycans known to be associated with human skin keratin and to skin keratin extracts. This binding of keratin extracts may explain initial bacterial attachment and clustering of the bacteria in the skin, ultimately leading to tissue destruction and ulceration caused by a cloud of secreted mycolactone at the site of infection.