Collagen

Collagen, the most abundant protein in vertebrates, is an extracellular, inextensible fibrous protein that comprises the major protein component of such stress-bearing structures as bones, tendons, and ligaments. The objective of this exercise is to develop an understanding of the fibrous portion of collagen and to show how the different levels of protein structure come together and form a highly ordered and stable fiber. Collagen's properties of rigidity and inextensibility are due to this highly ordered structure. The part of collagen without structural order is not illustrated in this model. This part of the protein complex having a different amino acid composition, lysine and hydroxylysine are particularly important residues, is globular in nature and not as structurally organized. Lysine and hydroxylysine form covalent crosslinks in the protein complex, thereby adding strength and some flexibility to the fiber. This covalent crosslinking continues throughout life and produces a more rigid collagen and brittle bones in older adults. Go to Collagen Structure & Function for information on functions and disorders of collagen, and a link to assembly movies of the triple helix of types I and IV is available in the External Links section.

Structure of a Segment

A fiber segment is made up of 5 tropocollagens, each is shown in a . One limitation of this model of collagen segment is that instead of having flush cut ends as shown here, the ends of the tropocollagen in an actual fiber section would be . When the tropocollagens come together to form the fiber segment, they actually overlap one another in a staggered pattern. The presents of these staggered ends permit the tropocollagens from different segments to associate, and this associations result in the formation of a strong fiber. Add tropocollagens at a time to form the fiber section, , , , . View fiber segment as . Viewing the segment from the end one can see that without the side chains being displayed the center of the fiber is empty. Each contains 3 parallel peptide chains wrapped around one another to make a right-handed triple helix that is 87 Å long and ~10 Å in diameter. Tropocollagen displayed as only. The peptides making up the tropocollagen have a conformation which produces a . This helix has a rise which gives 3.3 residues/turn compared to 3.6 for the α helix. The shows that the psi and phi angles of the collagen helix are different from the α-helix. The two clusters shown here are outside of the area expected for an α-helix. Review where you would expect a cluster of α-helix residues to be located.

Primary Structure and Higher Levels of Structure

of the peptide in wireframe display. Identify the amino acids making up the peptide by resting the cursor on a residue and observing the name in the label (Stopping spin will make this easier.). Which three amino acids are present in the peptide in a reocurring pattern? Collagen is characterized by a distinctive repeating sequence: (Gly-X-Y)n where X is often Pro, Y is usually 5-hydroxyproline (Hyp), and n may be >300. The model (4CLG) being studied contains this of residues - Gly-Pro-Hyp. This sequence produces in the peptide chain a with 3.3 residues per turn and a pitch (rise per turn) of 10.0 Å. Looking down the axis of a tropocollagen displayed as wireframe, glycine can be seen of the triple helix. Proline and the hydroxyproline are on the of the triple helix. With the hydroxyproline being on the outside of the triple helix, its hydroxy groups can be involved in hydrogen bond formation as will be seen in the next section. The primary structure of repeatingGly-Pro-Hyp determines these positions in the tropocollagen. The triple helix structure requires the close packing of the interior residues of the triple helix making only a small volume available for side chains in the interior of the tropocollagen. The consist of only a hydrogen (Realize that in this model the hydrogen on the α carbon is not displayed.), and therefore only glycine cam accommodate this close packing in the interior of the triple helix. , Gly:16 on each of three different chains, are close packed together. The gray atoms of the yellow and lime Gly are the α-carbons, and only a hydrogen could fit between the carbons and the atoms of the adjacent Gly. on each of the 3 chains are shown close packed to the three Gly (lime, cyan, yellow). Adding to the complex extends the close packing around the small glycines.

Maintainance Forces

Intra-tropocollagen Attractions

Intra-tropocollagen attractions are primarily hydrogen bonds formed between the peptides in the triple helix (tropocollagen). The three polypeptide chains are in position by one residue, that is, a Pro on Chain A is at the same level along the triple helix axis as a Gly on Chain B and a Hyp on Chain C. This staggered arrangement not only a Gly main chain NH (imino group) with a Pro main chain O (carbonyl oxygen) on one of the other peptides but also brings the two groups close enough to form hydrogen bonds between the imino hydrogen and the carbonyl oxygen. This alignment of Gly occurs with each of the three peptides so that the Gly hydrogen of (orange) to the Pro carbonyl oxygen on Chain B and likewise Gly of (yellow) Pro and Gly of (green) Pro. The force of these hydrogen bonds extending the length of the tropocollagen add up to a strong attractive force which mantain the integrity of the tropocollagen. Since the main chain N atoms of both Pro and Hyp residues lack H atoms, only Gly can provide hydrogen to form these hydrogen bonds.

Inter-tropocollagen Attractions

Hydrogen bonds are also an important inter-tropocollagen force which holds the tropocollagens together in the fiber segment. These hydrogen bonds are formed between the hydroxy hydrogen of a Hyp which, as see above, is a residue on the of the triple helix and a backbone carbonyl oxygen. The hydroxyl groups are the atoms that extend out the most from the center of the triple helical structure. As the peptides in a tropocollagen twist about each other they come into from peptides in adjacent tropocollagens. The two peptide highlighted in spacefill are located in two different tropocollagens. Notice that they make contact with each other in the middle of the strands, and a hydrogen bond is located at this point of contact. The consist of the oxygen of a carbonyl of a Hyp in a peptide of one tropocollagen and the hydroxyl hydrogen of a Hyp in a peptide of another tropocollagen. Another example shows from two different tropocollagens making contact at the ends of the fiber segment, and of course it is within these regions where the inter-tropocollagen attractions occur. At one end a is formed between a hydrogen of Hyp in one peptide and an oxygen of a Gly carbonyl in the second peptide. At the other end of the two peptides a donates its electrons to a Hyp hydroxyl hydrogen. Show the in the context of the six peptides of the two tropocollagens. The above examples of hydrogen bonding illustrate that Hyp plays a central role in maintaining the structures of both the tropocollagen and the collagen fiber.

Effect of a Mutation

PDB ID: 1cag is a synthetic model of whose peptides contain thirty residues and have a of (Pro-Hyp-Gly)4-Pro-Hyp-Ala-(Pro-Hyp-Gly)5 (Ala displayed as large wireframe and colored as C O N). Viewing 1CAG from the side of the fiber shows: the is only partially visible, and therefore is buried in the interior, being much more visible is positioned closer to the surface, being clearly visible is on the surface, and being a substitute for Gly is only partially visible. The of the tropocollagen is shown with the Ala appearing as olive and the Hyp close to the Ala appearing as dark brown. Notice that the surface at these Hyp protrudes more than it does at the other Hyp. This protrusion is due to the fact that the packing about the Ala side chains is not as close as it is about the Gly. Using 1CAG to compare the packing around Ala to that around Gly shows that the is not as close as it is about the Gly.