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October, 2006
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JoVE 10692

Ribosomes translate genetic information encoded by messenger RNA (mRNA) into proteins. Both prokaryotic and eukaryotic cells have ribosomes. Cells that synthesize large quantities of protein—such as secretory cells in the human pancreas—can contain millions of ribosomes.

Ribosomes are composed of ribosomal RNA (rRNA) and proteins. Ribosomes are not surrounded by a membrane (i.e., despite their specific cell function, they are not an organelle). In eukaryotes, rRNA is transcribed from genes in the nucleolus—a part of the nucleus that specializes in ribosome production. Within the nucleolus, rRNA is combined with proteins that are imported from the cytoplasm. The assembly produces two subunits of a ribosome—the large and small subunits. These subunits then leave the nucleus through pores in the nuclear envelope. Each one large and small subunit bind to each other once mRNA binds to a site on the small subunit at the start of the translation process. This step forms a functional ribosome. Ribosomes may assemble in the cytosol—called free ribosomes—or while attached to the outside of the nuclear envelope or endoplasmic reticulum—called bound ribosomes. Generally, free ribosomes synthesize proteins used in the cytoplasm, while bound ribosomes synthesize proteins that are inserted into membranes, packaged into org

 Core: Biology

ELISPOT Assay: Detection of IFN-γ Secreting Splenocytes

JoVE 10497

Source: Tonya J. Webb1
1 Department of Microbiology and Immunology, University of Maryland School of Medicine and the Marlene and Stewart Greenebaum Comprehensive Cancer Center, Baltimore, Maryland 21201

ELISPOT is a standardized, reproducible assay used to detect cellular immune responses. The assay utilizes an enzyme-linked…


Golgi Apparatus

JoVE 10970

As they leave the Endoplasmic Reticulum (ER), properly folded and assembled proteins are selectively packaged into vesicles. These vesicles are transported by microtubule-based motor proteins and fuse together to form vesicular tubular clusters, subsequently arriving at the Golgi apparatus, a eukaryotic endomembrane organelle that often has a distinctive ribbon-like appearance.

The Golgi apparatus is a major sorting and dispatch station for the products of the ER. Newly arriving vesicles enter the cis face of the Golgi—the side facing the ER—and are transported through a collection of pancake-shaped, membrane-enclosed cisternae. Each cisterna contains unique compositions of enzymes and performs specific protein modifications. As proteins progress through the cis Golgi network, some are phosphorylated and undergo removal of certain carbohydrate modifications that were added in the ER. Proteins then move through the medial cisterna, where they may be glycosylated to form glycoproteins. After modification in the trans cisterna, proteins are given tags that define their cellular destination. Depending on the molecular tags, proteins are packaged into vesicles and trafficked to particular cellular locations, including the lysosome and plasma membrane. Specific markers on the membranes of these vesicles allow them to dock

 Core: Biology

Adult Stem Cells

JoVE 10810

Stem cells are undifferentiated cells that divide and produce more stem cells or progenitor cells that differentiate into mature, specialized cell types. All the cells in the body are generated from stem cells in the early embryo, but small populations of stem cells are also present in many adult tissues including the bone marrow, brain, skin, and gut. These adult stem cells typically produce the various cell types found in that tissue—to replace cells that are damaged or to continuously renew the tissue. The epithelium lining the small intestine is continuously renewed by adult stem cells. It is the most rapidly replaced tissue in the human body, with most cells being replaced within 3-5 days. The intestinal epithelium consists of thousands of villi that protrude into the interior of the small intestine—increasing its surface area to aid in the absorption of nutrients. Intestinal stem cells are located at the base of invaginations called crypts that lie between the villi. They divide to produce new stem cells, as well as daughter cells (called transit amplifying cells) that divide rapidly, move up the villi and differentiate into all the cell types in the intestinal epithelium, including absorptive, goblet, enteroendocrine, and Paneth cells. These mature cells continue to move up the villi as they carry out their functions, except Paneth cell

 Core: Biology

Induced-fit Model

JoVE 10734

Most chemical reactions in cells require enzymes—biological catalysts that speed up the reaction without being used up or permanently changed. They work by reducing the activation energy needed for the reactants to be converted into the products. Enzymes, which are usually proteins, work by binding to a substrate—a reactant molecule that they act upon.

Enzymes exhibit substrate specificity, meaning that they can only bind to certain substrates. This is mainly determined by the shape and chemical characteristics of their active site—the region of the enzyme that binds to the substrate. According to the induced-fit model of enzyme activity, this binding changes the conformation—or shape—of both the enzyme and the substrate. This brings the substrate closer to the higher energy transition state needed for the reaction to occur, for instance, by weakening its bonds so that it can more readily react. Enzymes may also speed up a reaction by creating conditions within the active site that are more conducive for the reaction to proceed than the surrounding cellular environment. Once the products of the reaction are formed, they are released from the active site and the enzyme can be used to catalyze reactions once again.

 Core: Biology
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