Lysozyme C

Lysozyme is a cornerstone of innate immunity. The canonical mechanism for bacterial killing by lysozyme occurs through the hydrolysis of cell wall peptidoglycan (PG). Conventional type (c-type) lysozymes are also highly cationic and can kill certain bacteria independently of PG hydrolytic activity. Reflecting the ongoing arms race between host and invading microorganisms, both gram-positive and gram-negative bacteria have evolved mechanisms to thwart killing by lysozyme. In addition to its direct antimicrobial role, more recent evidence has shown that lysozyme modulates the host immune response to infection. The degradation and lysis of bacteria by lysozyme enhance the release of bacterial products, including PG, that activate pattern recognition receptors in host cells. Yet paradoxically, lysozyme is important for the resolution of inflammation at mucosal sites. This review will highlight recent advances in our understanding of the diverse mechanisms that bacteria use to protect themselves against lysozyme, the intriguing immunomodulatory function of lysozyme, and the relationship between these features in the context of infection.

Ever since lysozyme was discovered by Fleming in 1922, this protein has emerged as a model for investigations on protein structure and function. Over the years, several high-resolution structures have yielded a wealth of structural data on this protein. Extensive studies on folding of lysozyme have shown how different regions of this protein dynamically interact with one another. Data is also available from numerous biotechnological studies wherein lysozyme has been employed as a model protein for recovering active recombinant protein from inclusion bodies using small molecules like l-arginine. A variety of conditions have been developed in vitro to induce fibrillation in hen lysozyme. They include (a) acidic pH at elevated temperature, (b) concentrated solutions of ethanol, (c) moderate concentrations of guanidinium hydrochloride at moderate temperature, and (d) alkaline pH at room temperature. This review aims to bring together similarities and differences in aggregation mechanisms, morphology of aggregates, and related issues that arise using the different conditions mentioned above to improve our understanding. The alkaline pH condition (pH 12.2), discovered and studied extensively in our lab, shall receive special attention. More than a decade ago, it was revealed that mutations in human lysozyme can cause accumulation of large quantities of amyloid in liver, kidney, and other regions of gastrointestinal tract. Understanding the mechanism of lysozyme aggregation will probably have therapeutic implications for the treatment of systemic nonneuropathic amyloidosis. Numerous studies have begun to focus attention on inhibition of lysozyme aggregation using antibody or small molecules. The enzymatic activity of lysozyme presents a convenient handle to quantify the native population of lysozyme in a sample where aggregation has been inhibited. The rich information available on lysozyme coupled with the multiple conditions that have been successful in inducing/inhibiting its aggregation in vitro makes lysozyme an ideal model protein to investigate amyloidogenesis.

Lysozyme is relevant to the innate immune system as a vital protein for crustaceans. In the present study, we cloned and characterized a novel c-type lysozyme gene (LvLYZ) from the Pacific white shrimp (Litopenaeus vannamei). The obtained full-length cDNA of LvLYZ was 990 bp and contained an open reading frame of 693 bp. Its deduced amino acid sequence consisted of 230 amino acids (aa) with a 17 aa signal peptide at the N-terminal and 130 aa functional domains. The multiple sequence alignment (MSA) indicated that the typical active sites in LvLYZ were similarly conserved as c-type lysozymes from other species. The transcription of LvLYZ appeared in all detected tissues and had relatively higher expression levels in hemocytes, hepatopancreas, gill and intestine. The mRNA expression profiles of LvLYZ were up-regulated in hemocyte and hepatopancreas post the stimulation of Vibrio parahaemolyticus or white spot syndrome virus (WSSV), respectively. The recombinant protein of LvLYZ (rLvLYZ) exhibited antibacterial activities against various microbes, including Escherichia coli, Vibrio splendidus, Micrococcaus luteus, Vibrio parahaemolyticus and Staphylococcus aureus. These results indicated that LvLYZ could cope with bacteria in L. vannamei and may play a significant role in immune response against invading pathogens.