This project aimed to analyze protein lipidation, a widespread strategy for anchoring proteins to cellular membranes across all domains of life, in the model archaeon Haloferax volcanii. Using deletion mutants of aliA and aliB, the first archaeal enzymes shown to be involved in lipobox-containing protein biogenesis, we employed large-scale Triton X-114 fractionation followed by quantitative proteomics to systematically analyze AliA- and AliB-dependent lipoprotein lipidation. Deletion of aliA affected most predicted lipoproteins in Hfx. volcanii, markedly diminishing their TX-114 enrichment, indicating reduced hydrophobicity. This establishes AliA as the primary enzyme responsible for archaeal lipoprotein lipidation. In contrast, deletion of aliB affected only a small subset of predicted lipoproteins. In addition to defining distinct and non-redundant roles for AliA and AliB, this study provides the first large-scale experimental validation of predicted archaeal lipoproteins and identifies candidate components of the archaeal lipoprotein biogenesis pathway.