The 2014 Conference will bring together a diverse group of investigators that share a common interest in understanding how proteins cross or insert into biological membranes. Elucidation of protein transport mechanisms remains a fundamental objective of modern cell biology as 30% of all proteins are either transported across or integrated into cellular membranes. Achieving correct cellular compartmentalization by these mechanisms is essential for cell function, and different facets of protein translocation directly impact human health. The 2014 conference will be organized by research topic, each of which will incorporate talks employing different methodologies to solve a common conceptual problem. Research topics will include sorting and targeting of proteins among intracellular membranes, the passage of proteins through translocation channels, the mechanisms of membrane protein assembly, the structural biology of transport machines, specialized pathogenic transport pathways, biogenesis of organelles, and others. This Gordon Research Conference will foster the synthesis of new ideas by bringing together a combination of established leaders and early career scientists. In addition to invited speakers, short talks will be selected from submitted abstracts. The 2014 conference also features a pre-conference Gordon Research Seminar (GRS), organized by postdoctoral scientists, that will be oriented towards junior investigators. The aim is to provide participants with background on common mechanisms involved in protein transport, to discuss their ongoing research projects, and to facilitate interaction and networking with their peers within this scientific community. We fully anticipate that the talks, discussions, poster sessions, and other informal interactions at this GRC and GRS will contribute to advancing our understanding of molecular mechanisms of protein trafficking, which is essential to both prokaryotic and eukaryotic cells.